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Download The A HISTOMETRIC INVESTIGATION OF THE ACTIVITY OF THE PITUITARY-INTERRENAL AXIS IN JUVENILE COHO SALMON, ONCORHYNCHUS KISUTCH WALBAUM by DONALD J. McLEAY B.Sc. University of British Columbia, 1966 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS OF THE DEGREE OF DOCTOR OF PHILOSOPHY IN THE DEPARTMENT OF ZOOLOGY We accept this thesis as conforming to the required standard The University of British Columbia July, 1970 In presenting this thesis in partial fulfilment of the requirements for an advanced degree at the University of British Columbia, I agree that the Library shall make it freely available for reference and study. I further agree that permission for extensive copying of this thesis for scholarly purposes may be granted by the Head of my Department or by his representatives. It is understood that copying or publication of this thesis for financial gain shall not be allowed without my written permission. Department of Z^-O The University of British Columbia Vancouver 8, Canada ABSTRACT The activity of the pituitary-interrenal axis in juvenile coho salmon (Oncorhynchus kisutch) and the relationship of this activity to the numbers of circulating leucocytes has been studied, using histological and histometric techniques. To this end, the effects of injections of mammalian ACTH, of Cortisol acetate and of dexamethasone on the interrenal tissue, on the epsilon cells in the pituitary gland, and on the differential leucocyte counts were investigated. In addition, the activity of the pituitary-interrenal axis and the related changes in leucocyte counts were determined throughout their year of stream residence. Further, the response of this axis, along with corresponding hematological changes to environmental alterations in the laboratory, were studied. Injections of ACTH resulted in a dosage-related stimulation of the interrenal tissue of coho salmon fry. On the other hand, injections of Cortisol acetate and of dexamethasone produced a marked atrophy of the inter• renal tissue. All dosages of either ACTH or Cortisol acetate decreased the mean nuclear diameters of epsilon cells, and resulted in their degranulation. In addition, a decrease in numbers of circulating small lymphocytes and thrombocytes resulted from administration of all dosages of ACTH, Cortisol acetate or dexamethasone. It is proposed that the interrenal tissue of coho fry is capable of marked variations in activity, that this tissue is under pituitary control, and that a negative-feedback mechanism operates between the interrenal and the pituitary gland. Additionally, changes in pituitary-adrenocortical activity are reflected in characteristic alterations in numbers of certain circulating leucocytes. The pituitary-interrenal axis of juvenile coho salmon in their natural habitat is inactive, from the time of emergence in spring, and through summer and early fall, compared with the winter and spring samples ii of yearling coho. In addition, numbers of circulating small lymphocytes were decreased in the winter and spring samples of yearling coho compared with summer and autumn samples. It is suggested that the increased activity of the pituitary-interrenal axis along with changes in leucocyte counts observed in the winter sample of juvenile coho salmon are related to cold-temperature acclimation. Furthermore, it is proposed that the increased pituitary- adrenocortical activity noted in the latest of the three spring samples of yearling coho salmon is associated with the transformation from parr to smolt. The interrenal tissue of juvenile coho salmon maintained in continuous darkness, or exposed to a continuously flashing light for varying numbers of days, was generally more active than that of corresponding control fish subjected to a twelve hour photoperiod. Additionally, small-lymphocyte and thrombocyte counts for darkness-maintained and flash-exposed fish were lower than values for corresponding control samples. On the other hand, no consistent differences in activity of the interrenal tissue were found when fish maintained in continuous light for varying numbers of days and • corresponding control fish were compared. It was observed that the pituitary- interrenal axis of juvenile coho salmon was initially stimulated following transfer of the fish from holding tanks to an altered environment. In addition, the activity of the interrenal tissue was increased by exposure of these fish to cold water temperatures; this increased interrenal activity was accompanied by a lymphopenia. It is suggested that the pituitary-interrenal axis of juvenile coho salmon is involved in cold-temperature acclimation. It is concluded that the pituitary-interrenal axis of juvenile coho salmon undergoes marked fluctuations in activity as a result of environmental alterations within the laboratory. Furthermore, an increase in pituitary-adrenocortical activity during acclimatization is characteristically reflected in a decrease in number of.circulating small lymphocytes. iii TABLE OF CONTENTS Page ABSTRACT i LIST OF TABLES vi LIST OF FIGURES ix ACKNOWLEDGEMENTS xi i i GENERAL INTRODUCTION 1 SECTION I: PRESENCE OF A PITUITARY-INTERRENAL AXIS AND ITS RELATIONSHIP TO THE CIRCULATING LEUCOCYTES 4 INTRODUCTION .... , 4 MATERIAL AND METHODS .... 8 Experimental Animals 8 Description of Holding Tanks and Experimental Compartments ... 8 Experiments 11 Autopsy Procedure 11 Histological Techniques 13 Karyometry .. -15 Statistical Evaluation of Results 16 RESULTS 18 Structure of the Interrenal Gland of Control Fry 18 Structure of the Rostral Pars Distalis of Control Fry ........ 22 Description of the Peripheral Blood Cell-types of Coho Fry 23 The Effect of ACTH Injections on the Pituitary-interrenal .Axis 27 The Effects of Cortisol and Dexamethasone Injections on the Pituitary-interrenal Axis 37 Staining of Interrenal Cells of Saline, ACTH.and Cortisol-Injected Fry for RNA 40 Effect of ACTH on Blood Cell Counts 42 Effect of Cortisol and Dexamethasone on Blood Cell Counts .... 45 Correlation Coefficients of Variables Measured in the ACTH- Injection Experiments 48 Correlation Coefficients of Variables Measured in the Cortisol and Dexamethasone Experiments . 51 DISCUSSION.... ... 56 iv Page SECTION II: THE ACTIVITY OF THE PITUITARY-INTERRENAL AXIS AND RELATED HEMATOLOGICAL CHANGES IN JUVENILE COHO CAPTURED SEASONALLY FROM BERTRAND CREEK 75 INTRODUCTION 75 MATERIAL AND METHODS 77 RESULTS ....................... 78 Seasonal Histology of the Head Kidney and Interrenal Gland .. 78 Seasonal Histology of the Epsilon Cells in the Rostral Pars Distalis 87 Mean Blood Cell Counts for Seasonal Samples 89 Correlation Coefficients of Variables Measured in the Seasonal Samples . 92 DISCUSSION .'. 96 SECTION III: VARIATIONS IN INTERRENAL ACTIVITY AND RELATED HEMATOLOGICAL CHANGES IN JUVENILE COHO SALMON EXPOSED TO DISSILIMAR ENVIRONMENTS WITHIN THE LABORATORY 105 INTRODUCTION 105 MATERIAL AND METHODS 107 Continuous-Darkness Experiments ... 107 Continuous-Light Experiments 108 Flashing-Light Experiments ..... 110 Correlation Matrix of Variables Measured from All Experiments Ill RESULTS .......... ...... 112 A. Continuous-Darkness Experiments 112 Histology of the Head Kidney and Interrenal Gland 112 Mean Blood Cell Counts . 118 Correlation Coefficients of Variables Measured in the Continuous-Darkness Experiments . 121 B. Continuous-Light Experiments 123 Histology of the Head Kidney and Interrenal Gland 123 Mean Blood Cell Counts 129 Correlation Coefficients of Variables Measured in the Continuous-Light Experiments 132 V Page C. Flashing-Light Experiments 134 Histology of the Head Kidney and Interrenal Gland 134 Mean Blood Cell Counts .. 141 Correlation Coefficients of Variables Measured in the Flashing-Light Experiments 144 Correlation Coefficients of Variables Measured from All Experiments 146 DISCUSSION ... 150 SUMMARIES 162 SECTION I ...1.......................... 162 SECTION II 163 SECTION III .;. 165 APPENDIX I ^............. ^ v............. 168 APPENDIX II ........... 169 REFERENCES 177 vi LIST OF TABLES TABLE PAGE I Details of materials injected and dosages received . .. 12 II Effect of ACTH-injections on interrenal nuclear diameters, number of interrenal nuclei per defined area of interrenal tissue, and epsilon cell nuclear diameters of coho fry 30 III Effect of Cortisol acetate- and dexamethasone-injections on interrenal nuclear diameters, number of interrenal nuclei per defined area of interrenal tissue, and epsilon cell nuclear diameters of coho fry 31 IV The effect of ACTH-injections on blood cell counts of coho fry ... 43 V The effect of Cortisol acetate- and dexamethasone- injections on blood cell counts of coho fry 46 VI Correlation matrix of variables measured from coho fry involved in the ACTH experiment . 49 VII Correlation matrix of variables measured from coho fry involved in the Cortisol experiment ^2 VIII Correlation matrix of variables measured from coho fry involved in the dexamethasone experiment 53 IX Mean interrenal nuclear diameters, number of interrenal nuclei per defined area of interrenal tissue, and epsilon cell nuclear diameters of juvenile coho salmon sampled seasonally from Bertrand Creek 83 X Blood cell counts of juvenile coho salmon taken seasonally from Bertrand Creek ....... 90 XI Correlation matrix of variables measured from juvenile coho salmon taken seasonally from Bertrand Creek 93 • XII Mean interrenal nuclear diameters, number of interrenal nuclei per defined area of interrenal tissue, and epsilon cell nuclear diameters of juvenile coho salmon involved in the
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