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Biochemical studies on sheep ovarian follicles maintained in organ culture
THERIOGENOLOGY
BIOCHEMICAL
STUDIES ON SHEEP OVARIAN FOLLICLES MAINTAINED IN ORGAN CULTURE. R.F. Seamark, F. Amato, Susan.Hendrickson, Leila Mak and R.M. Moor, Department of Obstetrics and Gynaecology, University of Adelaide, South Australia 5000. The development of techniques for organ culture has made possible detailed biochemical studies on the response of individual ovarian In this study we report on the tissues- to gonadotrophic stimulation. effects of human chorionic gonadotrophin (hCG 20 iu ml-') on oxygen uptake, glucose utilization, lactate production and cellular content of adenine nucleotides of sheep follicles isolated and maintained under the conditions of culture desribed by Moor(l). The results, expressed as nmols per mg wet weight tissue per hr, (meanvalue 2 SEM, No. of observations in parentheses) are summarised below. 4 ! Period in culture (h) 24 0 Oxygen consumption:/ Entire follicles 560260 (9) 530f44 (12) +hCG Theta 1080+164(6) i +hCG Granulosa 52214 (6) +hCG Glucose utilization 1 8.5+0.7(G) Entire follicles [ 9.6+0.8(6) -hCG I Lactate production:1 Entire follicles ! 16.1+2.0(6) 14.2+3.0(6) +hCG Tissue ATPt2) 13502220 2130+161(7) Tissue ADP(3) 30+20 75533 c Tissue AMPc3) 20250 185+33 0.97 0.91 ECR**
422233 (13) 374230 (10) 8602140 (6) 13072221 (6)
l *
Energy charge ratio = ATP + $ADP l P c.05 for effect of hCG. ATP + ADP + AMP The study indicates that the tissues are well maintained in culture. The increase in lactate production observed with hCG coincides with a marked increase in progestin secretion, and a transfer of steroidcgenic It is tempting function from the theta interna to membrana granulosa. to speculate that the increase in glycolytic activity of the follicles is due to the granulosa cells as they assume their new role. References: 1.
2.
3.
Moor, R.M. (1973) Oestrogen production by individual follicles explanted from ovaries of sheep. J. Peprod. Fert. 32, 545-548 Stanley, P.E. and%lliams, S.G. (19691 Use of the liquid scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme. Analyt. Biochem z, 381-392. Parkinson, P.I. and Medley, E. (1972) In 'Liquid Scintillation Counting' Eds. Crook, M-A., Johnson, P. and Scales, B., Heyden & Son London and N.Y. p. 109-113.
DlECEMBER
1976
VOL. 6 NO. 6
603
BIOCHEMICAL
STUDIES ON SHEEP OVARIAN FOLLICLES MAINTAINED IN ORGAN CULTURE. R.F. Seamark, F. Amato, Susan.Hendrickson, Leila Mak and R.M. Moor, Department of Obstetrics and Gynaecology, University of Adelaide, South Australia 5000. The development of techniques for organ culture has made possible detailed biochemical studies on the response of individual ovarian In this study we report on the tissues- to gonadotrophic stimulation. effects of human chorionic gonadotrophin (hCG 20 iu ml-') on oxygen uptake, glucose utilization, lactate production and cellular content of adenine nucleotides of sheep follicles isolated and maintained under the conditions of culture desribed by Moor(l). The results, expressed as nmols per mg wet weight tissue per hr, (meanvalue 2 SEM, No. of observations in parentheses) are summarised below. 4 ! Period in culture (h) 24 0 Oxygen consumption:/ Entire follicles 560260 (9) 530f44 (12) +hCG Theta 1080+164(6) i +hCG Granulosa 52214 (6) +hCG Glucose utilization 1 8.5+0.7(G) Entire follicles [ 9.6+0.8(6) -hCG I Lactate production:1 Entire follicles ! 16.1+2.0(6) 14.2+3.0(6) +hCG Tissue ATPt2) 13502220 2130+161(7) Tissue ADP(3) 30+20 75533 c Tissue AMPc3) 20250 185+33 0.97 0.91 ECR**
422233 (13) 374230 (10) 8602140 (6) 13072221 (6)
l *
Energy charge ratio = ATP + $ADP l P c.05 for effect of hCG. ATP + ADP + AMP The study indicates that the tissues are well maintained in culture. The increase in lactate production observed with hCG coincides with a marked increase in progestin secretion, and a transfer of steroidcgenic It is tempting function from the theta interna to membrana granulosa. to speculate that the increase in glycolytic activity of the follicles is due to the granulosa cells as they assume their new role. References: 1.
2.
3.
Moor, R.M. (1973) Oestrogen production by individual follicles explanted from ovaries of sheep. J. Peprod. Fert. 32, 545-548 Stanley, P.E. and%lliams, S.G. (19691 Use of the liquid scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme. Analyt. Biochem z, 381-392. Parkinson, P.I. and Medley, E. (1972) In 'Liquid Scintillation Counting' Eds. Crook, M-A., Johnson, P. and Scales, B., Heyden & Son London and N.Y. p. 109-113.
DlECEMBER
1976
VOL. 6 NO. 6
603