ICES Journal of Marine Science (2011), 68(10), 2053–2058. doi:10.1093/icesjms/fsr144

Telomere length analysis in species: Metapenaeus macleayi, verreauxi, and

Rosamond M. Godwin1*, Stewart Frusher2, Steven S. Montgomery3, and Jennifer Ovenden1 1Molecular Fisheries Laboratory, Agri-Science Queensland, Department of Employment, Economic Development and Innovation, Level 3 Ritchie Building (64A C-Wing), Research Road, The University of Queensland, Brisbane 4067, Australia 2Tasmanian Aquaculture and Fisheries Institute, University of , Private Bag 49, Hobart 7001, Australia 3Industry and Investment NSW, Cronulla Fisheries Research Centre of Excellence, PO Box 21, Cronulla, NSW 2230 Australia

*Corresponding Author: tel: +61 7 3346 6518; fax: +61 7 3346 6501; e-mail: [email protected] Downloaded from https://academic.oup.com/icesjms/article/68/10/2053/611301 by guest on 29 September 2021 Godwin, R. M., Frusher, S., Montgomery, S. S., and Ovenden, J. 2011. Telomere length analysis in crustacean species: Metapenaeus macleayi, Sagmariasus verreauxi, and Jasus edwardsii. – ICES Journal of Marine Science, 68: 2053–2058. Received 1 March 2011; accepted 20 June 2011; advance access publication 15 September 2011.

Estimates of age and growth in have been historically problematic and presented significant challenges to researchers. Current techniques of age determination provide valuable data, but also suffer from disadvantages. Telomeric DNA has been proposed as an age biomarker because it shortens with age in some species. In this study, the feasibility of using telomere length (TL) to estimate age was examined in the school Metapenaeus macleayi and the spiny Sagmariasus verreauxi and Jasus edwardsii. Carapace length (CL) was used as a surrogate for age, and terminal restriction fragment assays were used to test the relationship between TL and size. Degradation of telomeric DNA with time during storage significantly influenced TL estimates, particularly for M. macleayi. TLs obtained from species in this study were 10–20 kb. No relationship between CL and TL was detected for any of the test species, and TL did not differ between male and female M. macleayi. TLs of J. edwardsii pueruli were unexpectedly shorter than those of J. edwardsii adults. The suitability of TL as an age biomarker in crustaceans may be limited, but further research is needed to elucidate telomere dynamics in these species with their different life histories and lifespans. Keywords: age biomarker, age estimation, Australia, crustacean, telomere length, TRF.

Introduction In many vertebrate somatic cells, telomeres shorten with each Accurate assessments of age are required for understanding mitotic cell division over the ’s life as a result of incomplete growth, maturity, reproduction, longevity, and mortality of replication of the DNA (Watson, 1972). This shortening has been in a wild population (Campana, 2001). They are basic correlated with age in mammals and some birds (Haussmann and considerations in stock assessment models for sustainable manage- Vleck, 2002; Haussmann et al., 2003; Vleck et al., 2003; Nakagawa ment of fisheries. et al., 2004), as well as in fish (Hatakeyama et al., 2008; Hartmann Estimating age and growth of crustaceans has been historically et al., 2009). It is also correlated with age in oysters and shell size in problematic, presenting significant challenges for researchers. abalone (RMG, unpublished data). Therefore, telomere length Methods used to estimate age or growth include tag-recapture, (TL) has been suggested as a potential age biomarker in animals size frequency distributions, size or weight measurements, and where it is difficult to determine age using conventional lipofuscin accumulation. Such methods provide valuable data methods (von Zglinicki and Martin-Ruiz, 2005; Haussmann and but can be inaccurate, subject to biases, costly, time-consuming, Mauck, 2008). The telomere repeat sequence of crustaceans is and deleterious to the animal (reviewed by Wahle and Fogarty, (TTAGG)n, which is common in many species 2006). Direct measures of size or weight are only weakly correlated (Klapper et al., 1998; Sahara et al., 1999; Lang et al., 2004; with age in crustaceans (Zheng et al., 1995; Uglem et al., 2005). Vı´tkova´ et al., 2005; Elmore et al., 2008). Detailed studies of TL Lipofuscin pigments, which accumulate with age in crustacean in crustaceans, however, are yet to be conducted. neural tissue, have yielded limited success, and the accuracy of In this study, the relationship between TL and age (as indicated the method is still under debate (Harvey et al., 2008; Sheehy, by size) is examined in three crustacean species with different life- 2008); accumulation of lipofuscin is not independent of environ- spans and life-history characteristics. The test species were school mental effects, particularly temperature (Wahle and Fogarty, , Metapenaeus macleayi, and two species of spiny , 2006). Therefore, the development of an alternative, low-cost, Sagmariasus verreauxi (formerly Jasus verreauxi) and Jasus non-lethal method to estimate age in crustaceans reliably would edwardsii. All three are commercially important and common in be a major improvement for fisheries management. Australian waters. School prawns are short lived (lifespan 12–18 Telomeres are the structures of highly repeated DNA sequences months; Montgomery et al., 2010) and endemic to estuarine and and their associated proteins that protect the ends of the linear inshore waters from southern Queensland to eastern . chromosomes from degradation and fusion (Blackburn, 1991). Juvenile and subadult prawns inhabit estuaries, generally near

# The State of Queensland (through the Department of Employment, Economic Development and Innovation), 2011. 2054 R. M. Godwin et al. beds of seagrass. Adults are found mainly in marine waters, but 3-year-old lobsters were originally caught off Port Hacking as also in small numbers in estuaries (Ruello, 1977; Coles and recently settled pueruli. These small lobsters were then grown in Greenwood, 1983). Sagmariasus verreauxi and J. edwardsii captivity for some 3 years before sampling. The sampled J. edwardsii have relatively long lifespans, 30 years for S. verreauxi comprised two broad size classes, pueruli (CL unknown) and adults (Montgomery et al., 2009), and potentially longer for J. edwardsii. (CL 80–120 mm). Sagmariasus verreauxi is distributed in warmer waters along the Lobsters were transported to the laboratory alive and eutha- east coast of Australia and the North Island of nased in ice slurry. Sampled tissues (Table 1) were dissected (Montgomery et al., 2009), and J. edwardsii widely around from the animals, placed in tubes, immediately snap-frozen in southern mainland Australia, Tasmania, and New Zealand liquid nitrogen, and then transferred to a 2808C freezer for long- (Booth, 2007). term storage. Lobsters such as S. verreauxi and J. edwardsii exhibit indetermi- nate growth patterns (Wahle and Fogarty, 2006), and despite its Downloaded from https://academic.oup.com/icesjms/article/68/10/2053/611301 by guest on 29 September 2021 shorter life, M. macleayi is also likely to exhibit indeterminate Extraction of total genomic DNA and TRF assays growth. Hence, the three species grow continuously throughout Genomic DNA of high molecular weight was extracted from their life and theoretically have no upper size limit. frozen tissue using standard methods (Sambrook and Russell, The aim of this study was to determine whether TL was suitable 2001). Extracted crustacean DNA appeared to be susceptible to as an age biomarker in crustaceans. If suitable, it could provide a degradation, so to minimize the co-purification of contaminating molecular method of age determination that would allow age to be material, a smaller quantity of fresh tissue was used during cell estimated from small samples of tissue, a non-lethal and cheaper lysis. Hence, 50 mg of tissue was digested per millilitre of digestion alternative to conventional methods of collecting age data. The buffer (10 mM Tris, pH 8.0, 5.0 mM EDTA, pH 8.0, 0.5% SDS, strategy was to compare TLs in at least two divergent age/size –1 200 mgml proteinase K). Different combinations of tempera- cohorts per species and to add data from other cohorts if ture and incubation period were optimal for cell lysis and digestion differences were indicated. TL can be estimated from very small of fresh tissue for each species (Table 2). Proteins and other con- quantities of tissue using PCR methods (Cawthorn, 2002), but taminants were removed by salt precipitation in 5 M potassium in the absence of any available sequence data for these test acetate and phenol/chloroform extractions. species, a terminal restriction fragment (TRF) assay was used in The resulting DNA was dissolved in TE (Tris–EDTA) buffer, this initial study. Our hypothesis was that there would be an pH 8.0, with 1 mM dithiothreitol added to minimize degradation inverse relationship between TL and age, as has been observed in and oxidation of the telomeric DNA. All DNA samples were stored many vertebrate species. at 48C. The quality and the quantity of genomic DNA were assessed by gel electrophoresis before each assay. High-quality Material and methods DNA samples contained molecular weight fragments .23 kb Sampling and tissue storage and a minimal presence of low molecular weight fragments Size (as indicated by carapace length, CL) was used as a surrogate (Figure 1). Degradation was evidenced by most of the DNA for age because wild specimens of unknown age were studied appearing as a band or smear of very low molecular weight. The (Table 1). Three broad size classes of M. macleayi, comprising exclusion of obviously degraded samples from TL assays was a approximately equal numbers of males and females, were analysed standard part of the experimental design. for TL. CLs were recorded before abdominal tissue from the tail TLs were estimated from the extracted DNA using a TRF assay, area was dissected from the exoskeleton for subsequent analysis. which was performed using a TeloTAGGG TL assay kit (Roche Sagmariasus verreauxi samples encompassed four general size Diagnostics Australia Pty Ltd, Castle Hill, New South Wales, classes, from pueruli to very large (CL 11–159 mm). The Australia), with minor modification (Table 2).

Table 1. Details of crustacean sample groups, the number in each group, and the available morphometric information. Size class (number/size Species Source class) CL (mm) Weight (g) Sampled tissues Metapenaeus Lake Wooloweyah, an estuarine lake near Yamba, Small (n ¼ 15) 10–14 Unknown Abdominal tissue macleayi New South Wales (29829′S 153818′E) minus exoskeleton Medium (n ¼ 16) 16–23 Unknown Abdominal tissue minus exoskeleton Yamba vicinity, New South Wales offshore Large (n ¼ 19) 16–25 Unknown Abdominal tissue (29826′S 153822′E) minus exoskeleton Sagmariasus Waters off Port Hacking, New South Wales (34817′S Puerulus, recently 11–15 0.6–1.4 Abdominal tissue verreauxi 151818′E), then grown at Cronulla Fisheries, NSW settled (n ¼ 6) minus exoskeleton Small, 3 years old 65–87 158–364 Muscle and gill (n ¼ 5) Coffs Harbour, New South Wales (30818′S 15389′E) Large (n ¼ 5) 103–130 670–1 150 Muscle and gill Coffs Harbour, New South Wales (30818′S 15389′E) Very large (n ¼ 6) 150–159 .1 800 Muscle and gill Jasus edwardsii Eastern Tasmanian waters (41852′S 148818′E) Puerulus, recently Unknown Unknown Abdominal tissue settled (n ¼ 7) minus exoskeleton Sandstone Bluff area, Tasmania (42820′S 148813′E) Adult (n ¼ 9) 80–120 Unknown Muscle Telomere length analysis in crustaceans 2055

Table 2. Summary of modifications to the TeloTAGGG TL assay kit for successful TRF assays on crustacean species. Cell-lysis Restriction Species conditions endonucleases Electrophoresis Gel Metapenaeus 508C for 3 h HinfI and RsaI378C Conventional 35V for 20 h at 48C 0.8% agarose in macleayi for 3 h 1 × TAE buffer Sagmariasus 378C AluI and HpyCH4V at Fluctuating inverted-field gel electrophoresis, with the 1.0 % agarose in verreauxi overnight 378C for 3 h programme optimized for 25–50 kb fragments (Fwd 180 V, 0.5 × TBE buffer Rev 120 V, switch time 0.4–0.8 linear shape) Recirculating buffer, with run time 20 h Jasus edwardsii 508C for 3 h AluI and HpyCH4V at Conventional 35 V for 20 h at 48C 0.8% agarose in 378C for 5 h 1 × TAE buffer Downloaded from https://academic.oup.com/icesjms/article/68/10/2053/611301 by guest on 29 September 2021 Total-intensity data inside each box and the molecular weight at the centre of each box were estimated using Quantity One software (Biorad). The background subtraction method and the formula for mean TRF calculation were as per the Roche protocol.

Data analysis For school prawns, the main effects of size (CL) and sex on TL were investigated. Equal numbers of male and female prawns were used in each of three size categories. The effect of DNA degra- dation during storage time on TL was also investigated during the data analysis. Storage time was the number of days that extracted DNA was stored at 48C before TRF assay. GenStat (Payne et al., 2009) was used to fit general linear models (GLMs; McCullagh and Nelder, 1989) for TL. DNA storage time and CL were continu- Figure 1. High-quality uncut genomic DNA from 13 female and 12 ous variates, and sex was a factor. Residual plots indicated that the male medium-sized M. macleayi subsequently used for TL assay. normal model with identity link was acceptable. Splines and Molecular weight markers (bp) are shown on the left side. polynomials of the variates were considered, with the latter (with degree 2) adopted for interpretation. All interactions were Although complete digestion of M. macleayi DNA was obtained trialled, but dropped from the final model, because they were using the restriction endonucleases HinfI and RsaI from the Roche not significant (p . 0.05). kit, preliminary experiments revealed only partial digestion of S. For the two spiny lobsters, GLMs were also fitted to datasets for verreauxi and J. edwardsii DNA. Therefore, a range of other fre- each species using GenStat (Payne et al., 2009). Individual animals quently cutting endonucleases, such as AciI, AluI, HpyCH4V, were the independent experimental units, with split plots adopted MseI, TaqI, and Tsp5091, was tested in various combinations on for the replicate measurements of the same animals. TL was the those species. The most efficient combination of enzymes for the dependent variable, and size or CL the independent factor for lobster species was AluI and HpyCH4V. Genomic DNA (2 mg the analyses. Inspection of the residual plots indicated that no per sample) was digested with the appropriate enzyme combi- transformation of the data was necessary. Post hoc unpaired nation for 3–5 h at 508C. t-tests were used to compare the data, which could not be included Telomere smears of S. verreauxi were not efficiently in the same GLM as a result of aliasing. resolved using conventional agarose gel electrophoresis, so fluctuating inverted-field gel electrophoresis (Biorad, Australia) was employed. A mixture of two DNA size markers was used (l Monocut mix, plus l HindIII; New England BioLabs Results Inc., USA), allowing size estimation of DNA fragments up to Metapenaeus macleayi 50 kb. Denatured DNA was transferred to nylon membranes, and the There was no difference in TL between sexes in M. macleayi ¼ ¼ telomeric DNA was detected with the Roche TeloTAGGG kit (F1,176 1.49; p 0.224), nor was there any relationship ′ ¼ ¼ except where a 3 digoxigenin end-labelled (TTAGG) probe between TL and CL (F2,176 1.59; p 0.207). Mean TLs for 6 + (Sigma Genosys Pty Ltd, Castle Hill, Australia) was used. TRF female and male prawns were 10 020 250 bp (s.e.) and + assays were repeated three times on each set of samples, with 9709 215 bp (s.e.), respectively. samples loaded in random order on each gel. Degradation of telomeric DNA during storage significantly affected the TL measurements in school prawns. Values of TL decreased by 30% as storage time increased from 1 to 3 weeks Determination of TL (F2,176¼ 35.55; p , 0.001; Figure 2). Adjusted values in the Mean TRF measurements were determined using the Roche graph are the observed values adjusted for other terms in the TeloTTAGG TL assay protocol from images captured on X-ray model. This reduction in TL took place despite controlling for films. Each lane was overlaid with a grid of 25 equal-sized boxes. genomic DNA quality immediately before the TRF assay. 2056 R. M. Godwin et al.

Table 3. Mean and s.e. for TL estimates from different tissues and size classes of S. verreauxi. Tissue TL (bp; mean + s.e.) Puerulus tail (abdomen exoskeleton) 17 908 + 366 Adult gill 17 603 + 565 Adult muscle 16 318 + 486 Adult gill muscle, combined mean 16 961 + 1 131 Adult size class Small (3 years old) 16 720 + 618 Large 18 190 + 785 Very large 15 961 + 630 Downloaded from https://academic.oup.com/icesjms/article/68/10/2053/611301 by guest on 29 September 2021

Figure 2. The effect of DNA storage time (between extraction and TRF analysis, in days) on TL values (in bp) of M. macleayi.

Sagmariasus verreauxi TLs of S. verreauxi were close to 16–18 kb, some twice the length of that of M. macleayi, and varying little between the sampled tissues (Table 3). Among adult lobsters, there was no difference between gill and muscle tissue (F1,94¼ 2.89; p ¼ 0.092). Additionally, the combined mean TL for adult gill and muscle tissue did not differ from the TL of puerulus abdominal tissue (t ¼ 0.80; p ¼ 0.43). No relationship between size and TL was detected among the 3-year-old, large, and very large S. verreauxi (F ¼ 2.14; 2,93 Figure 3. The effect of DNA storage time (between extraction and p ¼ 0.124; Table 3). Data from pueruli were excluded from this TRF analysis, in days) on TL values (in bp) of S. verreauxi. comparison because they were based on a mixture of abdominal tissues. TLs of older lobsters were based on single tissues, either gill or muscle. When CL was used as a more-precise measure of size, and the relationship between TL and CL tested by linear p , 0.001). The apparent lengthening of J. edwardsii telomeres as regression, the result was also not significant (F1,94¼ 0.99; animals grow (age) is not conclusive, because the results are con- p ¼ 0.323). The coefficient of variation (CV) in TL among founded by the different tissues used for each group of animals. sampled pueruli was 11.6% and 21.2% among adult individuals. The result here may therefore be solely an effect of the presence Like M. macleayi, the duration of DNA storage time signifi- of non-muscle tissues in pueruli samples. cantly affected TL estimates of S. verreauxi (F1,94¼ 9.11; In adult J. edwardsii, there was no relationship between CL and p ¼ 0.003; Figure 3). Several S. verreauxi DNA extracts had been TL (F1,7¼ 0.37; p ¼ 0.563). A CV of 10.0% was recorded among stored for nearly 10 months before TRF assay and, although the individual pueruli and 4.8% among adults for the TL. The vari- quality of the DNA was checked immediately before assay, there ation in TL among pueruli of both lobster species was comparable, was an average decline in TL of 24.8% over 300 d. The degradation but the variation among adult S. verreauxi was four times greater observed for S. verreauxi was slower than observed for M. macleayi, than that among adult J. edwardsii, perhaps reflecting greater however, for which there was a similar degree of degradation in variability in TL in the S. verreauxi population. However, this one-tenth of the time. could be an artefact caused by the degradation of some of the S. verreauxi samples. Jasus edwardsii The number of days for which the DNA had been stored TL assays were conducted on J. edwardsii using DNA from the before TRF assay did not affect the TL estimates of J. edwardsii abdominal tissue of pueruli and the muscle tissue of adults. Gill significantly (F1,17¼ 0.03; p ¼ 0.868). The susceptibility of J. tissue was unavailable for assaying. TLs of J. edwardsii were inter- edwardsii DNA to degradation cannot, however, be ruled mediate between those of M. macleayi and S. verreauxi. out because there was only 10 d difference between the shortest The average TL of pueruli (10 259 bp + s.e. 139) was unexpectedly and the longest storage times of any DNA extracts (18 vs. shorter than for adult J. edwardsii (15 124 bp + s.e. 164; t ¼ 22.6; 28 d). Telomere length analysis in crustaceans 2057

Discussion We expected TL to decrease with age in a short-lived crustacean Methodological considerations such as M. macleayi, but this was not the case. The age of the wild- caught specimens was unknown, but we can predict from the life The results of this study have highlighted the susceptibility of crus- cycle of M. macleayi that small estuarine individuals would have tacean telomeric DNA to degradation during storage and its been younger than large ocean prawns. This is because juveniles impact on the accuracy of TL estimations. This effect was inhabit estuaries, whereas adults are mainly found in open seas most pronounced for M. macleayi, for which TL estimates were (Ruello, 1977; Coles and Greenwood, 1983). The absence of up to 30% lower if DNA had been stored for a few weeks. telomere shortening with age in M. macleayi suggests that Metapenaeus macleayi DNA was mainly from muscle tissue, but telomere-maintenance mechanisms are present in both short- digestive tract tissues could also have been present, contributing and long-lived crustaceans. Additionally, such mechanisms do to the problems of degradation. In previous studies, we found not seem to vary between the sexes of M. macleayi. that the DNA from digestive tissue of marine organisms such as High telomerase expression has been detected in all differen- oysters is highly susceptible to degradation (data not shown). Downloaded from https://academic.oup.com/icesjms/article/68/10/2053/611301 by guest on 29 September 2021 tiated tissues of the green C. maenas, which has a lifespan Takubo et al. (2002) also stressed the need to check for DNA of 5–6 years (Elmore et al., 2008), and both reproductive and degradation caused by autolysis and artificial changes, in a somatic tissues of P. japonicus (Lang et al., 2004). Elmore et al. variety of human tissues. They checked DNA samples before (2008) found telomerase activity at significant levels in a variety TRF assay using pulsed field electrophoresis and found that a con- of tissues of both long- and short-lived aquatic species of diverse siderable number of samples degraded and had to be omitted from taxa. Moreover, they suggested that these high levels of telomerase their study. expression are not related to the longevity of aquatic animals, but Perhaps telomeric DNA degrades at a faster rate than non- to their ability to regenerate tissues after injury. telomeric DNA in stored DNA samples. The use of in-gel visualiza- Future research using the TRAP assay (Kim et al., 1994) could tion to check DNA quality was not sensitive enough to detect this usefully assess the relevance of telomerase expression to the main- specific type of degradation. Steps taken in this study to prevent tenance of TL in crustaceans. For example, telomerase activity may the degradation of telomeric DNA during extraction and storage be correlated across species, with varying abilities to regenerate minimized, but never completely eliminated, the problem. limbs. Species with low telomerase activity may exhibit TL attri- Therefore, if accurate estimates of TL are to be made, assays tion with age. need to be conducted as soon as possible after crustacean DNA For the species examined in this study, there was no association has been isolated. Degradation of telomeric DNA during storage between TL and size. However, larval and juvenile stages were not is an important consideration for telomere researchers, and it examined, and those periods yield significant transition and may has the potential to contribute to the variability in published account for the fastest growth in the life history of a crustacean. data. Further research, with more samples over a controlled It may also be the period of the most dramatic change in TL. period, would be necessary to quantify the degradation of stored For example, differences in TL have been found in different life telomeric DNA and should be a necessary first step when stages of the jellyfish Cassiopea andromeda. The bell region of working with crustaceans. the medusa had significantly longer telomeres than did polyps (Ojimi and Hidaka, 2010). Future research comparing early larva, metamorphic, juvenile, and adult life stages may help to elu- Relationship between TL and age cidate the mechanisms of crustacean telomere dynamics. The TLs for crustacean species (10–20 kb) in this study were Genetic and environmental effects may have contributed to the similar to those reported for other aquatic species, spanning a variability in TL within each species. Each group of animals was variety of taxa (Elmore et al., 2008). Although the telomere sourced from different places, and in some cases, the exact location repeat sequence has been identified in many crustaceans including of sampling was unknown (Table 1). The variability in the sampled lobster Homarus americanus (Klapper et al., 1998), green crab groups may therefore have limited our ability to detect any associ- Carcinus maenas (Elmore et al., 2008), and the prawns Penaeus ation between TL and age. Larger sample sizes from more con- japonicus (Lang et al., 2004) and P. semisulcatus (Okazaki et al., trolled sample groups are obviously needed to facilitate a 1993), there have been no detailed studies of TL in crustaceans. more-detailed analysis of telomere dynamics and age in There was no relationship between TL and CL for the three crustaceans. crustacean species included in this study, despite the different lifespans. This was anticipated for the two lobster species Acknowledgements because telomerase, the enzyme involved in telomere repair and We thank Majorie D’Sousa, Vivek Mitter, and Raewyn Street for maintenance, is very active in all fully differentiated tissues of technical assistance during the project and David Mayer for help the American clawed lobster, H. americanus (Klapper et al., with the statistical analyses. Also, thanks are due to the project 1998). Lobsters have a high regenerative potential, exhibit indeter- steering committee members for their support and advice: minate growth, negligible senescence, and low rates of tumour for- Damien Broderick, Ian Brown, Melissa Brown, Phillip Gaffney, mation. It has been suggested that sustained telomerase expression John Russell, and Brian Patterson. The project was co-funded by is necessary to facilitate the continued growth and replacement of the Australian Fisheries Research and Development Corporation tissues across the whole life of lobsters (Klapper et al., 1998; Vogt, (project number 2007/033). 2008; Gomes et al., 2010). The lack of telomere shortening for S. verreauxi, and the apparent telomere lengthening for J. edwardsii between pueruli and very large adults, suggests that telomere- References maintenance mechanisms function throughout the life of both Blackburn, E. H. 1991. Structure and function of telomeres. Nature, these species. 350: 569–573. 2058 R. M. Godwin et al.

Booth, J. D. 2007. Jasus species. In Lobsters: Biology, Management, describe growth in a metapenaeid from waters off Australia. Aquaculture and Fisheries, pp. 340–358. Ed. by B. F. Phillips. Marine and Freshwater Research, 61: 1435–1445. Blackwell Publishing, Oxford, UK. 506 pp. Nakagawa, S., Gemmell, N. J., and Burke, T. 2004. Measuring ver- Campana, S. E. 2001. Accuracy, precision and quality control in age tebrate telomeres: applications and limitations. Molecular determination, including a review of the use and abuse of age vali- Ecology, 13: 2523–2533. dation methods. Journal of Fish Biology, 59: 197–242. Ojimi, M., and Hidaka, M. 2010. Comparison of telomere length Cawthorn, R. M. 2002. Telomere measurement by quantitative PCR. among different life cycle stages of the jellyfish Cassiopea Nucleic Acids Research, 30: 1–6. andromeda. Marine Biology, 157: 2279–2287. Coles, R., and Greenwood, J. 1983. Seasonal movement and size distri- Okazaki, S., Tsuchida, K., Maekawa, H., Ishikawa, H., and Fujiwara, H. bution of three commercially important Australian prawn species 1993. Identification of a pentanucleotide telomeric sequence, (Crustacea: ) within an estuarine system. Marine and (TTAGG)n, in the silkworm Bombyx mori and in other insects. Freshwater Research, 34: 727–743. Molecular and Cellular Biology, 13: 1424–1432. Elmore, L. W., Norris, M. W., Sircar, S., Bright, A. T., McChesney, P.

Payne, R. W., Harding, S. A., Murray, D. A., Baird, D. B., and Soutar, Downloaded from https://academic.oup.com/icesjms/article/68/10/2053/611301 by guest on 29 September 2021 A., Winn, R. N., and Holt, S. E. 2008. Upregulation of telomerase D. M. 2009. GenStat for Windows, 12 edn. Introduction. VSN function during tissue regeneration. Experimental Biology and International, Hemel Hempstead. 256 pp. Medicine, 233: 958–967. Ruello, N. V. 1977. Migration and stock studies on the Australian Gomes, N. M. V., Shay, J. W., and Wright, W. E. 2010. Telomere school prawn Metapenaeus macleayi. Marine Biology, 41: 185–190. biology in Metazoa. FEBS Letters, 584: 3741–3751. Sahara, K., Marec, F., and Traut, W. 1999. TTAGG telomeric repeats in Hartmann, N., Reichwald, K., Lechel, A., Graf, M., Kirschner, J., Dorn, chromosomes of some insects and other . Chromosome A., Terzibasi, E., et al. 2009. Telomeres shorten while Tert Research, 7: 449–460. expression increases during ageing of the short-lived fish Sambrook, J., and Russell, D. W. 2001. Molecular Cloning: a Nothobranchius furzeri. Mechanisms of Ageing and Development, Laboratory Manual, part 3. Cold Spring Harbor Laboratory 130: 290–296. Press, Cold Spring Harbor, pp. B.9; E.3–E.4. Harvey, H. R., Secor, D. H., and Ju, S. J. 2008. The use of extractable lipofuscin for age determination of crustaceans: Reply to Sheehy Sheehy, M. R. J. 2008. Questioning the use of biochemical extraction to (2008). Marine Ecology Progress Series, 353: 307–311. measure lipofuscin for age determination of : Comment on Ju et al. (1999, 2001). Marine Ecology Progress Series, 353: 303–306. Hatakeyama, H., Nakamura, K-I., Izumiyama-Shimomura, N., Ishii, A., Tsuchida, S., Takubo, K., and Ishikawa, N. 2008. The teleost Takubo, K., Izumiyama-Shimomura, N., Honma, N., Sawabe, M., Oryzias latipes shows telomere shortening with age despite con- Arai, T., Kato, M., Oshimura, M., et al. 2002. Telomere lengths siderable telomerase activity throughout life. Mechanisms of are characteristic in each human individual. Experimental Ageing and Development, 129: 550–557. Gerontology, 37: 523–531. Haussmann, M. F., and Mauck, R. A. 2008. New strategies for Uglem, I., Belchier, M., and Sva˚sand, T. 2005. Age determination of telomere-based age estimation. Molecular Ecology Resources, 8: European lobsters ( L.) by histological quanti- 264–274. fication of lipofuscin. Journal of Crustacean Biology, 25: 95–99. Haussmann, M. F., and Vleck, C. M. 2002. Telomere length provides a Vı´tkova´, M., Kra´l, J., Traut, W., Zrzavy´, J., and Marec, F. 2005. The new technique for aging animals. Oecologia, 130: 325–328. evolutionary origin of insect telomeric repeats, (TTAGG)N. Haussmann, M. F., Vleck, C. M., and Nisbet, I. C. T. 2003. Calibrating Chromosome Research, 13: 145–156. the telomere clock in common terns, Sterna hirundo. Experimental Vleck, C. M., Haussmann, M. F., and Vleck, D. 2003. The natural Gerontology, 38: 787–789. history of telomeres: tools for aging animals and exploring the Kim, N. W., Piatyszek, M. A., Prowse, K. R., Harley, C. B., West, M. D., aging process. Experimental Gerontology, 38: 791–795. Ho, P. L. C., Coviello, G. M., et al. 1994. Specific association of Vogt, G. 2008. How to minimize formation and growth of tumours: human telomerase activity with immortal cells and cancer. potential benefits of decapod crustaceans for cancer research. Science, 266: 2011–2015. International Journal of Cancer, 123: 2727–2734. Klapper, W., Ku¨hne, K., Singh, K. K., Heidorn, K., Parwaresch, R., and von Zglinicki, T., and Martin-Ruiz, C. M. 2005. Telomeres as bio- Krupp, G. 1998. Longevity of lobsters is linked to ubiquitous telo- markers for ageing and age-related diseases. Current Molecular merase expression. FEBS Letters, 439: 143–146. Medicine, 5: 197–203. Lang, G. -H., Wang, Y., Nomura, N., and Matsumura, M. 2004. Wahle, R. A., and Fogarty, M. J. 2006. Growth and development: Detection of telomerase activity in tissues and primary cultured understanding and modelling growth variability in lobsters. In lymphoid cells of Penaeus japonicus. Marine Biotechnology, 6: Lobsters: Biology, Management, Aquaculture and Fisheries, pp. 347–354. 1–43. Ed. by B. F. Phillips. Blackwell Publishing, Oxford, UK. McCullagh, P., and Nelder, J. A. 1989. Generalized Linear Models, 2nd 506 pp. edn. Chapman and Hall, London. 511 pp. Watson, J. 1972. Origin of concatemeric T7 DNA. Nature New Montgomery, S. S., Liggins, G. W., Craig, J. R., and McLeod, J. R. 2009. Biology, 239: 197–201. Growth of the Jasus verreauxi (: Zheng, J., Murphy, M. C., and Kruse, G. H. 1995. A length-based Palinuridae) off the east coast of Australia. New Zealand Journal population-model and stock–recruitment relationships for red of Marine and Freshwater Research, 43: 113–123. king crab Paralithodes camtschaticus in Bristol Bay Alaska. Montgomery, S. S., Walsh, C. T., Haddon, M., Kesby, C. L., and Canadian Journal of Fisheries and Aquatic Sciences, 52: Johnson, D. D. 2010. Using length data in the Schnute model to 1229–1246.