DOCSLIB.ORG

Abnormal Spermatogenesis in RXR[ Mutant Mice

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Abnormal Spermatogenesis in RXR[ Mutant Mice Downloaded from genesdev.cshlp.org on September 23, 2021 - Published by Cold Spring Harbor Laboratory Press Abnormal spermatogenesis in RXR[ mutant mice Philippe Kastner, t Manuel Mark, ~ Mark Leid, 2 Anne Gansmuller, William Chin, 3 Jesus M. Grondona, Didier D6cimo, Wojciech Krezel, Andr6e Dierich, and Pierre Chambon 4 Institut de G6n4tique et de Biologie Mol6culaire et Cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS)/Institut National de la Sant6 et de la Recherche M6dicale (INSERM)/Universit6 Louis Pasteur (ULP1, Collhge de France, BP 163-67404 ILLKIRCH-CEDEX, C.U. de Strasbourg, France We have generated mouse lines in which the RXRI3 gene was disrupted by homologous recombination. Approximately 50% of the RXR~ homozygous mutants died before or at birth, but those that survived appeared normal except that the males were sterile, owing to oligo-astheno-teratozoospermia. Failure of spermatid release occurred within the germinal epithelium, and the epididymis contained very few spermatozoa that, in addition, exhibited abnormal acrosomes and tails. There was a progressive accumulation of lipids within the mutant Sertoli cells, which were histochemically characterized as unsaturated triglycerides. In old mutant males, progressive degeneration of the germinal epithelium occurred, ending with the formation of acellular lipid-filled tubules. The selective expression of RXR[~ in Sertoli cells, together with the timing of appearence of the histological abnormalities, suggests that the primary defect resulting from the mutation resides in these cells. [Key Words: Sertoli cells; lipid metabolism defect; male sterility; seminiferous epithelium degeneration; retinoid X receptor] Received August 21, 1995; revised version accepted October 31, 1995. The three retinoid X receptors, RXRa, RXRB, and RXR~, receptor (VDR), and the peroxisome proliferator-acti- are members of the vertebrate nuclear receptor super- vated receptors (PPARs) (for review, see Mangelsdorf et family (Leid et al. 1992a, b; Mangelsdorf et al. 1992, 1994; al. 1992; Liu and Linney 1993; Doll4 et al. 1994; Nagata Chambon 1994; Giguhre 1994; Glass 1994; Kastner et al. et al. 1994; Desvergne and Wahli 1995). Finally, RXRs 1994b). In the mouse, RXRa and RXRB transcripts ap- may also be the heterodimeric partners of a number of pear to be widely expressed in the embryo and in adult orphan nuclear receptors, including members of the tissues, whereas the distribution of RXR-¢ transcripts is FXR/RLD-1/LXR/UR family (Apfel et al. 1994; Song et more restricted (Mangelsdorf et al. 1992; Liu and Linney al. 1994; Forman et al. 1995a; Seol et al. 1995; Teboul et 1993; Doll6 et al. 1994; Nagata et al. 1994). In vitro al. 1995; Willy et al. 1995; note that UR has also been DNA-binding studies and studies in transfected cells named OR-1 or RIP15), members of the NGFI-B/ cultured in vitro have provided evidence indicating that NURR-1 family (Forman et al. 1995b; Perlmann and RXRs could be involved in several signaling pathways. Jansson 1995), and the orphan receptor MB67 (Baes et al. First, as 9-cis retinoic acid (RA) (9C-RA)-dependent tran- 1994). scriptional regulators, RXR homodimers may transduce Whether RXRs engaged in heterodimeric associations some of the effects of the active retinoid derivatives of act as 9C-RA-dependent transcriptional regulators to vitamin A. Second, RXRs may also play a role in the synergistically control initiation of transcription or are retinoid signaling pathway, as heterodimeric partners for transcriptionally silent, simply allowing their partners the retinoic acid receptors (RARoL, RARf3, and RARe/) to bind efficiently to the response elements of their cog- that act as all-trans RA (T-RA)- or 9C-RA-dependent nate target genes, is largely unknown. Several studies transcriptional regulators. Third, RXRs may be involved performed in vitro and in transfected cultured cells have as heterodimeric partners in additional signaling path- suggested that the 9C-RA-dependent transcriptional ac- ways mediated by other nuclear receptors, which include tivity of the RXR partner is dependent on both the nature the thyroid hormone receptors (TRs), the vitamin D3 and the ligand occupancy of the heterodimeric partner, as well as on the nature of the bound response element (Durand et al. 1994; Kurokawa et al. 1994; Forman et al. tThese authors contributed equally. 1995b; Perlmann and Jansson 1995; Willy et al. 1995). Present addresses: 2College of Pharmacy,Oregon State University, Cor- Interestingly, Roy et al. (1995) have shown recently that, vallis, Oregon 97331 USA; SHarvardMedical School, Dept. of Medicine, Boston, Massachusetts 02115 USA. in P19 and F9 embryonal carcinoma cells, RAR- and 4Corresponding author. RXR-specific ligands have synergistic effects to activate 80 GENES& DEVELOPMENT 10:80-92 © 1996 by Cold Spring Harbor LaboratoryPress ISSN 0890-9369/96 $5.00 Downloaded from genesdev.cshlp.org on September 23, 2021 - Published by Cold Spring Harbor Laboratory Press Male sterility in RXR~ mutant mice endogenous RA target genes and to induce differentia- A BC (~ Probes I I , ,~ I--I 5' 3" tion, which indicates that, at least in some cases of RXR/ w-r Sp K XN?.B,.. KE B B,.S ep.Nh NhBB SpKX j .,, , , i ,(, RAR partnerships, both receptors can be implicated in RXR~ locus (+) ' transcriptional activation of target genes. In contrast to the wealth of information related to the targeting construct ~I , lkb 3' " 5' functions of RXRs in vitro, very little is known to date PGK-NEO(A +) concerning the actual physiological role of RXRs in vivo. K XN.': _=B=..I__~_EHB Sp ..'-"BB ,Sp.== BB, Sp,r" KX , Notably, it is unknown whether, as a receptor for 9C- mutant locus (-) S p RA, RXR transduces some of the multiple effects of ret- • 5 "1 inoids. In this respect, it is noteworthy that most of the (+)1 9kb I | Spel digest I J probe A abnormalities found in vitamin A-deficient fetuses or 7kb adult mice are reproduced in mice bearing mutations in (+) I~-~-~-I ] BamHI one or several RARs, which demonstrate that RARs are (-) ~1 digest 1kb probeB involved in the physiological transduction of the RA sig- nal (Lohnes et al. 1993, 1994; Lufkin et al. 1993; Men- delsohn et al. 1994). The possible function of RXRR has ''+' @ © +~i" +'~'~ been investigated by examining homozygous null mice .•&' generated by targeted disruption of the RXRa gene in -v 3kb embryonic stem (ES) cells (Kastner et al. 1994a; Sucov et probe A probe B Spe I 7kb BamHI al. 1994). These mutants exhibit fetal heart and eye de- 0-- lkb fects similar to those occurring in the fetal vitamin A-de- ficiency (VAD) syndrome, thus suggesting that RXR~ t 2 3 / 3kb mediates some of the effects of vitamin A. Moreover, P robe C BamHI [ these studies have revealed a strong synergy between mutations in RAR and RXR genes for the generation of 1234567 abnormalities that were either less severe in single RXR or RAR mutants or not present at all. It was also shown Figure 1. The RXR[3 mutation. (al Targeting the RXRff gene. A that some specific RXR/RAR pairs are much more effi- map of the wild-type RXRI3 gene is shown at the top. (.) Exons cient than others at generating some of the abnormali- (numbering as in Nagata et al. 1994). Probe A is a NotI-KpnI ties seen in the fetal VAD syndrome (Kastner et al. fragment; probe B is a 500-bp BamHI-HindIII fragment imme- 1994a; P. Kastner, M. Mark, and P. Chambon, unpubl.). diately upstream of the deleted region; probe C is the HindIII- EcoRI fragment (corresponding to the deleted region). (Sp) SpeI; Taken together, these data strongly suggest that RAR/ (K) KpnI; (N) NotI; (B) BamHI; (E) EcoRI; (Nh) NheI; (X)XhoI; RXR heterodimers are the major functional units respon- (H) HindIII. (b) Targeted ES cells. Southern blot analysis of SpeI- sible for transducing the retinoid signal during develop- restricted DNA from wild type (WT) and the HA67 and HA9 ES ment. cell clones, analyzed with the 5' probe A. (c) Southern blot anal- We now report the generation of RXR~ null mutant ysis of mutant mice. (Top) The analysis of a litter using BamHI- mice. Approximately 50% of these mutants die in utero restricted DNA and probe B. (Bottom) Corresponds to an iden- or very shortly after birth for unknown reasons. Surpris- tical blot hybridized with probe C. ingly, the remaining null mutants are externally indis- tinguishable from their wild-type or heterozygous litter- mates. However, RXR~-/- males are sterile, owing to abnormal germ cell maturation, leading to oligo-as- HA9) transmitted the mutation, thus generating two theno-teratozoospermia. These sperm abnormalities lines of mutant mice. For both lines, crosses between most probably reflect an indispensable function of RXR~ heterozygotes (RXRS +/-) yielded viable homozygote in the Sertoli cells, which in prepubertal mutant males mutants (RXR~-/-), which were externally indistin- start to accumulate lipid droplets. The possible involve- guishable from their wild-type and heterozygote litter- ment of PPARs in the generation of the testis abnormal- mates. Homozygote females were fertile, but males were ities seen in RXR~ null mutants is discussed. sterile (see below). As expected, a probe spanning the genomic region en- coding the RXR~ DNA-binding domain did not detect any hybridizing fragment in homozygotes {probe C in Results Fig. lc). Therefore, even though RXRI3 exons 1, 2, and 5-9 are still present in the genome of homozygotes, a RXR[3 mutant mice functional DNA-binding RXR~ protein cannot be pro- The RXR~ gene was disrupted by homologous recombi- duced in the mutants. RT-PCR analysis of total RNA nation in ES cells with a replacement-type vector in isolated from several tissues showed that transcripts cor- which the genomic sequences encoding the DNA-bind- responding to exon 2, as well as transcripts containing ing domain (3' region of exons 3 and 4) were replaced exons 5-9, were present at a low level in RXR~ - / - mice with a PGK-NEO(A +) cassette (Fig.
Load more

Recommended publications
  • Identification of Differentially Expressed Genes of Primary
    Cell Research (2004); 14(6):507-512 ARTICLE http://www.cell-research.com Identification of differentially expressed genes of primary spermatocyte against round spermatid isolated from human testis using the laser capture microdissection technique Gang LIANG1,4, Xiao Dong ZHANG1, Lu Jing WANG1, Yu Shen SHA2, Jian Chao ZHANG2, Shi Ying MIAO1, Shu Dong ZONG2, Lin Fang WANG1,*, S.S. KOIDE3 1National Laboratory Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, 5 Dong Dan San Tiao, 100005 Beijing, China 2National Research Institute for Family Planning, WHO Collaboration Center for Research in Human Reproduction, Beijing, 12 Da Hui Si, 100081 Beijing, China 3Center for Biomedical Research, Population Council, 1230 York Avenue, New York, NY 10021, USA 4Chinese National Human Genome Center, Beijing, 3-707 North Yong Chang Road BDA, Beijing 100176, China ABSTRACT The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differen- tiation and development. In the present study, over 10,000 primary spermatocytes and round spermatid cells were successfully isolated by LCM. Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed. The average insert size of the cDNA isolated from 75 randomly picked white clones was 500 bp, ranging from 250 bp to 1.7 kb. Using the dot-blot method, a total of 421 clones were examined, resulting in the identification of 390 positive clones emitting strong signals.
    [Show full text]
  • Kidney, Renal Tubule – Dilation
    Kidney, Renal Tubule – Dilation Figure Legend: Figure 1 Kidney, Renal tubule - Dilation in a male B6C3F1 mouse from a chronic study. Dilated tubules are noted as tracts running through the cortex and outer medulla. Figure 2 Kidney, Renal tubule - Dilation in a male F344/N rat from a chronic study. Tubule dilation is present throughout the outer stripe of the outer medulla, extending into the cortex. Figure 3 Kidney, Renal tubule - Dilation in a male B6C3F1 mouse from a chronic study. Slight tubule dilation is associated with degeneration and necrosis. Figure 4 Kidney, Renal tubule - Dilation in a male F344/N rat from a chronic study. Tubule dilation is associated with chronic progressive nephropathy. Comment: Renal tubule dilation may occur anywhere along the nephron or collecting duct system. It may occur in focal areas or as tracts running along the entire length of kidney sections (Figure 1). 1 Kidney, Renal Tubule – Dilation Renal tubule dilation may occur from xenobiotic administration, secondary mechanisms, or an unknown pathogenesis (see Kidney – Nephropathy, Obstructive (Figure 2). Dilation may result from direct toxic injury to the tubule epithelium interfering with absorption and secretion (Figure 3). It may also occur secondary to renal ischemia or from prolonged diuresis related to drug administration. Secondary mechanisms of tubule dilation may result from lower urinary tract obstruction, the deposition of tubule crystals, interstitial inflammation and/or fibrosis, and chronic progressive nephropathy (Figure 4). A few dilated tubules may be regarded as normal histologic variation. Recommendation: Renal tubule dilation should be diagnosed and given a severity grade. The location of tubule dilation should be included in the diagnosis as a site modifier.
    [Show full text]
  • Anatomical and Morphological Study of the Kidneys of the Breeding Emu (Dromaius Novaehollandiae)
    Turkish Journal of Zoology Turk J Zool (2016) 40: 314-319 http://journals.tubitak.gov.tr/zoology/ © TÜBİTAK Research Article doi:10.3906/zoo-1506-21 Anatomical and morphological study of the kidneys of the breeding emu (Dromaius novaehollandiae) 1, 2 3 2 3 Katarzyna MICHAŁEK *, Danuta SZCZERBIŃSKA , Marta GRABOWSKA , Danuta MAJEWSKA , Maria LASZCZYŃSKA 1 Department of Physiology, Cytobiology, and Proteomics, Faculty of Biotechnology and Animal Husbandry, West Pomeranian University of Technology in Szczecin, Szczecin, Poland 2 Department of Poultry and Ornamental Bird Breeding, West Pomeranian University of Technology in Szczecin, Szczecin, Poland 3 Department of Histology and Developmental Biology, Pomeranian Medical University, Szczecin, Poland Received: 15.06.2015 Accepted/Published Online: 05.01.2016 Final Version: 07.04.2016 Abstract: We analyzed 16 kidneys obtained from 15-year-old emus, Dromaius novaehollandiae. Emus were kept at an experimental farm in Poland. The results showed that each kidney was composed of 3 parts: cranial, medial, and caudal divisions. Histological results demonstrated that the kidneys consisted of 2 zones: the cortex and the medulla. The cortex made up the majority of the kidney, while the medulla formed only a small portion of the organ. Proximal and distal tubules and 2 types of glomeruli (looped and loopless) were localized in the cortex. Each of these glomeruli was characterized by tightly arranged mesangial cells. Proximal and distal tubules had a distinctive simple low cuboidal epithelium. The luminal surface of the proximal tubules had a brush border membrane, formed by numerous microvilli. The renal medulla of emu kidneys formed irregularly positioned characteristic cones of different sizes.
    [Show full text]
  • Skates and Rays Diversity, Exploration and Conservation – Case-Study of the Thornback Ray, Raja Clavata
    UNIVERSIDADE DE LISBOA FACULDADE DE CIÊNCIAS DEPARTAMENTO DE BIOLOGIA ANIMAL SKATES AND RAYS DIVERSITY, EXPLORATION AND CONSERVATION – CASE-STUDY OF THE THORNBACK RAY, RAJA CLAVATA Bárbara Marques Serra Pereira Doutoramento em Ciências do Mar 2010 UNIVERSIDADE DE LISBOA FACULDADE DE CIÊNCIAS DEPARTAMENTO DE BIOLOGIA ANIMAL SKATES AND RAYS DIVERSITY, EXPLORATION AND CONSERVATION – CASE-STUDY OF THE THORNBACK RAY, RAJA CLAVATA Bárbara Marques Serra Pereira Tese orientada por Professor Auxiliar com Agregação Leonel Serrano Gordo e Investigadora Auxiliar Ivone Figueiredo Doutoramento em Ciências do Mar 2010 The research reported in this thesis was carried out at the Instituto de Investigação das Pescas e do Mar (IPIMAR - INRB), Unidade de Recursos Marinhos e Sustentabilidade. This research was funded by Fundação para a Ciência e a Tecnologia (FCT) through a PhD grant (SFRH/BD/23777/2005) and the research project EU Data Collection/DCR (PNAB). Skates and rays diversity, exploration and conservation | Table of Contents Table of Contents List of Figures ............................................................................................................................. i List of Tables ............................................................................................................................. v List of Abbreviations ............................................................................................................. viii Agradecimentos ........................................................................................................................
    [Show full text]
  • Anatomia Associada Ao Comportamento Reprodutivo De
    Jimena García Rodríguez Anatomia associada ao comportamento reprodutivo de Cubozoa Anatomy associated with the reproductive behavior of Cubozoa São Paulo 2015 Jimena García Rodríguez Anatomia associada ao comportamento reprodutivo de Cubozoa Anatomy associated with the reproductive behavior of Cubozoa Dissertação apresentada ao Instituto de Biociências da Universidade de São Paulo para obtenção de Título de Mestre em Ciências, na Área de Zoologia Orientador: Prof. Dr. Antonio Carlos Marques São Paulo 2015 García Rodríguez, Jimena Anatomia associada ao comportamento reprodutivo de Cubozoa 96 páginas Dissertação (Mestrado) - Instituto de Biociências da Universidade de São Paulo. Departamento de Zoologia. 1. Cubozoa; 2. Histologia; 3. Reprodução. I. Universidade de São Paulo. Instituto de Biociências. Departamento de Zoologia. Comissão Julgadora Prof(a) Dr(a) Prof(a) Dr(a) Prof. Dr. Antonio Carlos Marques A mis padres, hermana y en especial a mi abuelita “Caminante, son tus huellas el camino y nada más; Caminante, no hay camino, se hace camino al andar. Al andar se hace el camino, y al volver la vista atrás se ve la senda que nunca se ha de volver a pisar. Caminante no hay camino sino estelas en la mar” Antonio Machado, 1912 Agradecimentos Em primeiro lugar, eu gostaria de agradecer ao meu orientador Antonio Carlos Marques, Tim, pela confiança desde o primeiro dia, pela ajuda tanto pessoal como profissional durante os dois anos de mestrado, pelas discussões de cada tema tratado e estudado e pelas orientações que tornaram possível a elaboração deste trabalho. Agradeço também o apoio institucional do Instituto de Biociências e do Centro de Biologia Marinha da Universidade de São Paulo.
    [Show full text]
  • Spermatogenesis in Vitro
    SPERMATOGENESIS IN VITRO INDUCTION OF PROLIFERATION, MEIOSIS AND DIFFERENTIATION Mário Sousa Lab Cell Biology Institute of Biomedical Sciences (ICBAS) University of Porto [email protected] Spermatogonia A SPERMATOGENESIS IN VITRO Preleptotene Pachytene spermatocytes spermatocytes 26 days Spermatogonia B 16 days Elongated spermatids 2-3 days Secondary spermatocytes 7-11 days 5-8 days 2-3 days 2-3 days Elongating Round spermatids spermatids 16 days 16 days OBJECTIVES culture medium for long term cultures and cell differentiation cell and molecular processes at each germ cell stage germ cell lines homologous transplantation in vitro gene therapy 15 anejaculation cases M1 AB C D E Normal karyotypes Absence of Y microdeletions 600bp Conserved spermatogenesis SY254 (c) SY134 (b) SY142 (b) Mechanical dissociation SY152 (c) Erythrocyte lysis Enzymatic digestion Cell isolation by micromanipulation M2 Cell culture: SY14 (SRY) - Yp 5 CM SY84 (a) 5 CM + rFSH (25 U/L) SY157 (c) 5 rFSH + T (2 µmol/L) SY142 (b) Plated cells: 250 S + 100 SGA + 1000 ST1 + 100 ST2 Multiplex-PCR AZF a,b,c Yq11.2 Each testicle biopsy was collected in sperm preparation medium (SPM; Medicult, Copenhagen, Denmark) and squeezed with surgical blades. The resultant fluid was diluted with SPM and washed by centrifuging at 1,000 rpm (500-600 g), 2 times 5 minutes. The pellet was resuspended for 5 min in 2 ml of erythrocyte-lysing buffer (Verheyen et al., 1995), prepared with 155 mM NH4Cl, 10 mM KHCO3, and 2 mM EDTA in water, pH 7.2 with KOH (all from Sigma, Barcelone, Spain, cell culture tested), and filtered by 0.2 µm.
    [Show full text]
  • Determination of the Elongate Spermatid\P=N-\Sertolicell Ratio in Various Mammals
    Determination of the elongate spermatid\p=n-\Sertolicell ratio in various mammals L. D. Russell and R. N. Peterson Department of Physiology, School of Medicine, Southern Illinois University, Carbondale, IL 62901, U.S.A. Summary. Criteria were devised for determining the elongate spermatid\p=n-\Sertolicell ratio in various mammalian species at the electron microscope level. When data from particular species were pooled, the values were: rabbit, 12\m=.\17:1,hamster, 10\m=.\75:1; gerbil, 10\m=.\64:1;rat, 10\m=.\32:1; guinea-pig, 10\m=.\10:1;vole, 9\m=.\75:1;and monkey, 5\m=.\94:1. The elongate spermatid\p=n-\Sertolicell ratio is a measure of the workload of the Sertoli cell and is a prime factor determining their efficiency. The higher the ratio, the higher the sperm output is likely to be per given weight of seminiferous tubule parenchyma for a particular species. Introduction The number of spermatozoa provided in the ejaculate is determined by a number of factors but the major influence is the number of spermatozoa produced in the testis. In mammals that breed continuously testicular sperm production appears to be related to the size of the testis, especially the seminiferous tubule compartment. Here the kinetics of spermatogenesis dictate how many germ cells (spermatogonia) become committed to the spermatogenic process and also the time it takes these germ cells to go through various cell divisions and transformations to become a spermatozoon. The index of sperm production, or the daily sperm production, is expressed as the number of spermatozoa produced per day by the two testes of an individual, whereas the index of efficiency of sperm production is the number of spermatozoa produced per unit weight or volume of testicular tissue (Amann, 1970).
    [Show full text]
  • Nomina Histologica Veterinaria, First Edition
    NOMINA HISTOLOGICA VETERINARIA Submitted by the International Committee on Veterinary Histological Nomenclature (ICVHN) to the World Association of Veterinary Anatomists Published on the website of the World Association of Veterinary Anatomists www.wava-amav.org 2017 CONTENTS Introduction i Principles of term construction in N.H.V. iii Cytologia – Cytology 1 Textus epithelialis – Epithelial tissue 10 Textus connectivus – Connective tissue 13 Sanguis et Lympha – Blood and Lymph 17 Textus muscularis – Muscle tissue 19 Textus nervosus – Nerve tissue 20 Splanchnologia – Viscera 23 Systema digestorium – Digestive system 24 Systema respiratorium – Respiratory system 32 Systema urinarium – Urinary system 35 Organa genitalia masculina – Male genital system 38 Organa genitalia feminina – Female genital system 42 Systema endocrinum – Endocrine system 45 Systema cardiovasculare et lymphaticum [Angiologia] – Cardiovascular and lymphatic system 47 Systema nervosum – Nervous system 52 Receptores sensorii et Organa sensuum – Sensory receptors and Sense organs 58 Integumentum – Integument 64 INTRODUCTION The preparations leading to the publication of the present first edition of the Nomina Histologica Veterinaria has a long history spanning more than 50 years. Under the auspices of the World Association of Veterinary Anatomists (W.A.V.A.), the International Committee on Veterinary Anatomical Nomenclature (I.C.V.A.N.) appointed in Giessen, 1965, a Subcommittee on Histology and Embryology which started a working relation with the Subcommittee on Histology of the former International Anatomical Nomenclature Committee. In Mexico City, 1971, this Subcommittee presented a document entitled Nomina Histologica Veterinaria: A Working Draft as a basis for the continued work of the newly-appointed Subcommittee on Histological Nomenclature. This resulted in the editing of the Nomina Histologica Veterinaria: A Working Draft II (Toulouse, 1974), followed by preparations for publication of a Nomina Histologica Veterinaria.
    [Show full text]
  • Simplified and Objective Assessment of Spermatogenesis in Spinal Cord Injured Men by Flow Cytometry Analysis
    Paraplegia 31 (1993) 785-792 © 1993 International Medical Society of Paraplegia Simplified and objective assessment of spermatogenesis in spinal cord injured men by flow cytometry analysis I H Hirsch MD,! D Kulp-Hugues MD,! J Sedor MS,! P McCue MD, 2 M B Chancellor MD,! WEStaas MD,3 Deparments of 1 Urology, 2 Pathology, 3 Physical Medicine and Rehabilitation, Regional Spinal Cord Injury Center of the Delaware Valley, Jefferson Medical College, Philadelphia, PA, USA. Deterioration of the germinal epithelium of the testis is a known sequela of spinal cord injury (SCI) that may influence the outcome of male reproductive rehabilitation efforts. Quantitative testicular biopsy, currently regarded as the standard of assessing the integrity of spermatogenesis, has not gained wide­ spread clinical use because of its invasive nature and relative technical complex­ ity. Alternatively, aspiration DNA flow cytometry analysis of the testis has offered a potential method of spermatogenic assessment that meets both the requirements of simplicity and objectivity. The objective of this study is to determine the capability of flow cytometry to assess spermatogenesis following SCI. Eleven SCI men underwent incisional testicular biopsy with the specimen simultaneously submitted for quantitative evaluation of the germinal epithelium by both quantitative histometry and DNA flow cytometry. The haploid percen­ tage of cells showed highly significant levels of correlation with key micrometric parameters of the quantitative testicular biopsy: spermatid/tubule (p < 0.002) and the spermatid/Sertoli cell ratio (p < 0.0005). Since tissue procurement is accomplished less invasively for flow cytometry analysis, we recommend this method as the modality of assuring integrity of the germinal epithelium in candidates for reproductive rehabilitation.
    [Show full text]
  • Cytology and Kinetics of Spermatogenesis in the Rabbit
    CYTOLOGY AND KINETICS OF SPERMATOGENESIS IN THE RABBIT E. E. SWIERSTRA and R. H. FOOTE Department of Animal Husbandry, Cornell University, Ithaca, New York, U.S.A. {Received 21st August 1962) Summary. The cycle of the seminiferous epithelium of the rabbit was divided into eight stages, using as criteria the shape of the spermatid nucleus, the location of the spermatids and spermatozoa in regard to the basement membrane, the presence of meiotic figures and the release of spermatozoa from the lumen. The relative duration (frequency) of Stages 1 to 8 were 27-7, 13-4, 7-3, 11-0, 4-1, 15-7, 12-2 and 8-6%, respectively. Each stem cell (Type A spermatogonium) divided to produce two Type A spermatogonia. One of these was the starting cell for the next genera¬ tion, while the other gave rise to two intermediate-type spermatogonia. Three more spermatogonial divisions followed, producing sixteen primary spermatocytes from one Type A spermatogonium, as is characteristic for the bull and the ram, but unlike the rat, mouse and hamster. It was estimated that only 3-1 spermatids were generated from one primary spermatocyte, suggesting that in the rabbit there is considerable degeneration of spermatogenic cells during the two maturation divisions. INTRODUCTION Since the end of the last century, it has been known that well-defined cellular associations succeed one another in time in any one area of the semini¬ ferous tubules, and that along the tubules a more or less regular pattern of cell populations exists (Brown, 1885; Benda, 1887; von Ebner, 1888). This succession of cellular associations at any one location in the seminiferous tubules led to the concept of the cycle of the seminiferous epithelium defined by Leblond & Clermont (1952b) as that "series of changes occurring in a given area of the seminiferous epithelium between two successive appearances of the same cellular association".
    [Show full text]
  • On the Morphology of the Renal Tubules of Vertebrates
    AUTHOR'S ABSTRACT OF THIS PAPER ISSUED BY THE BIBLIOQRAPHIC SERVICE NOVEMBER 10. ON THE MORPHOLOGY OF THE RENAL TUBULES OF VERTEBRATES G. CARL HUBER Department of Anatomy, University of Michigaa TWENTY-TWO FIGURES A comparative study of the renal tubules of the different classes of vertebrates was projected nearly twenty years ago, soon after publishing results of a study on the development and shape of the uriniferous tubules of the higher mammah.1 In this study, as projected, it was purposed to reconstruct the excre- tory tubules of the pronephros and mesonephros of certain of the lower vertebrates, including amphibia, and the metanephric tubules of certain reptiles, birds and mammalia. In the re- construction, by the Born wax plate method, of the pronephric tubules of a larval toad, and mesonephric tubules of an adult frog and the metanephric tubules of certain reptilia no great difficulty was experienced, and such reconstructions were made, somewhat over fifteen years ago in conjunction with Professor Ward J. MacNeal, sometime Instructor in this department. On attempting to reconstruct the metanephric tubules of birds, it was learned after unsuccessful trials that this was beyond the limits of the method, so also with endeavors to reconstruct the metanephric tubules of adult mammalia. The projected study, therefore, was abandoned for a time. The form of the adult, mammalian renal tubule was later ascertained by specially devised methods of teasing.? This special method of teasing has relatively recently been successfully used in an investigation of the form of the metanephric tubule of birds. I am in a position now, therefore, to present figures giving the morphology 1 Huber, G.
    [Show full text]
  • Respiration and Elimination of Nitrogenous Wastes Forms and Functions of Plants and Animals
    MODULE - 2 Respiration and Elimination of Nitrogenous Wastes Forms and Functions of Plants and animals 14 Notes RESPIRATION AND ELIMINATION OF NITROGENOUS WASTES Every living organism needs energy to perform various life activities, and the process of respiration fulfils this energy requirement. You have already learnt in the lesson on food and nutrition that animals take in high energy organic molecules in the form of food. During respiration, this food is broken down in the presence of oxygen and energy is released during respiration. Respiration also produces carbon dioxide, a toxic substance which is eliminated from the body. Thus, uptake of oxygen and removal of carbon dioxide is an essential requirement of all animals. At the same time numerous other toxic wastes such as ammonia, and urea are also produced in the tissues during various cellular activities. Such toxic wastes need to be removed from the body. In this lesson you will learn about removal of nitrogenous wastes and maintenance of water and salt balance in the body. OBJECTIVES After completing this lesson you will be able to : z define respiration, breathing, inspiration, expiration and vital capacity; z describe briefly the gaseous exchange in earthworm and cockroach; z describe the parts of respiratory system in the human body and mention their functions; z draw a labeled diagram of human respiratory system; z differentiate between breathing and respiration; and inspiration and expiration; z describe the mechanism of breathing and its regulation; z describe the exchange
    [Show full text]