Neurophysin pathways in the normal and hypophysectomized rat

Summary The hypothalamo-extrahypophyseal neurophysin pathways (HEH) and the three hypothalamic nuclei secreting neurophysins, the supraoptic (SON), paraventricular (PVN) and suprachiasmatic (SCN) nuclei, of normal and hypophysectomized rats were studied by application of the immunoperoxidase procedure. Eight well-defined HEH pathways were recognized. Their main sites of projection were: lateral septum and subfornical organ (1 and 2); tractus diagonalis (3); medial nucleus of the amygdala and lateral ventricle (4); nucleus periventricularis thalami, nucleus habenulae lateralis and periaqueductal gray (5); periaqueductal gray, pineal organ, collicular recess and subependymal region of the fourth ventricle (6); dorsomedial nucleus and premammillary area (7); perimammillary region, corpus trapezoideum, ventral surface of medulla oblongata, nucleus tractus solitarii, nucleus commissuralis, substantia gelatinosa and formatio reticularis lateralis of the medulla oblongata and spinal cord (8).Neurophysin fibers of unknown origin were found in the frontal cerebral cortex.It was noted that in pathway 5 the amount of immunostainable material undergoes changes with age.The three neurophysin-secreting nuclei reacted differently following hypophysectomy. Among the HEH pathways the only one that seemed to be affected by hypophysectomy was that innervating the lateral septum.It is suggested that the neurons that survive hypophysectomy either do not project to the neural lobe or, alternatively, display axon collaterals projecting outside the neural lobe. Such a neuronal population could be the origin of the HEH pathways.Supported by Grant RSM-80-13 from the Directión de Investigaciones, Universidad Austral de Chile.The authors wish to acknowledge the valuable help of Mrs. Elizabeth Santibá~nez and Mr. Genaro Alvial.     

Effects on extrahepatic UDP-glucuronosyltransferases in hypophysectomized rat

The effects of hypophysectomy on hepatic and extrahepatic UDP-glucuronosyltransferase activities in adult male rats were observed. UDP-glucuronosyltransferase activities toward 1-naphthol decreased to 20-30% of control in the liver, kidney, lung, and testis. The mRNA of UGT1A6, which is an isoform contributing to the glucuronidation of various phenolic xenobiotics such as 1-naphthol, were decreased drastically in the liver, kidney, and testis by hypophysectomy. However, while bilirubin UDP-glucuronosyltransferase activity in the liver intensified, there was only a slight increase in the activity in the kidney and no alteration in the lung. The mRNA of UGT1A1, which is an isoform contributing to the glucuronidation of bilirubin, increased significantly in the liver and slightly in the kidney after hypophysectomy. These inductions and reductions in enzymatic activities and mRNA levels in each tissue were restored to control levels by intermittent injections of rat growth hormone. Interestingly, while hepatic UGT activity toward bisphenol A remained constant in hypophysectomized rats, the testicular UGT activity declined to 10-15% of control but returned to normal levels following growth hormone treatment, suggesting that an unknown UGT isoform (s) mediates bisphenol A glucuronidation in the testis. These results indicate that the expression of extrahepatic UGT is isoform-specific and regulated differentially in tissues by the pituitary gland.     

Regeneration of the supraoptico-hypophyseal and paraventriculo-hypophyseal tracts in the hypophysectomized rat

Summary The effect of hypophysectomy on the nerve fiber pattern in the median eminence and infundibular stem of the rat has been investigated by a slightly modified Bodian technique. Postoperative changes in the distribution of neurosecretory material and connective tissue and changes in vascularity have also been studied.Extensive regeneration of the fibers of the supraoptico-hypophyseal and paraventriculo-hypophyseal tract could be demonstrated. It is most pronounced at the distal extremity of the infundibular stem but occurs also in the rostral part of the infundibular stem and in the median eminence. Regeneration starts in the second postoperative week and is completed about four weeks later. The nervous regeneration observed in the pituitary area after hypophysectomy is more extensive than is usually encountered after lesions elsewhere in the central nervous system.It could be demonstrated moreover that neurosecretory material accumulates at the same sites in which the terminals of the regenerated nerve fibers can be found. Hypophysectomy also causes an increase in capillary density and connective tissue content of the infundibular stem. Accumulations of neurosecretory material are always found in areas showing a high capillary density and a considerable amount of connective tissue.Factors which might be responsible for the extensive nervous regeneration in the pituitary area are discussed as are the factors determining the pattern of outgrowth of the regenerating nerve fibers. Morphological aspects of storage of posterior pituitary hormones are considered in the light of data in the literature and the results of the present work.Partly supported by a U.S. Public Health grant to Dr. E. Scharrer and a travel grant to the author from the Netherlands Organization for pure scientific Research (ZWO).     

The estrogen 2-hydroxylase activity of the gonadotropin-stimulated hypophysectomized immature rat ovary

Using high-performance liquid chromatography and a combination of electrochemical and radiometric flow detection for 2-[14C]hydroxyestradiol, changes in estrogen 2-hydroxylase activity in the microsomal fraction of rat ovarian homogenates were followed. Injection of human chorionic gonadotropin (hCG) at 12-hr intervals to hypophysectomized immature rats stimulated hypertrophy of the theca-interstitial tissue and produced a profound increase in enzyme activity. With the last injection of hCG at 96 hr the peak serum concentration of hCG was reached 12 hr later and then decreased exponentially with a half-time of 13 hr. However, enzyme activity remained elevated for at least 60 hr before beginning to fall. Pregnant mare's serum gonadotropin (PMSG) also produced an increase in activity, which was apparently limited to the thecal-interstitial tissue because freshly removed granulosa cells from the mature follicles had undetectable activity levels. Administration of anti-PMSG antiserum after enzyme activity had been increased resulted in a prompt fall in activity, as did injection of hCG to mimic an ovulatory surge of LH. The results indicate that the thecal-interstitial tissue of the rat ovary has estrogen 2-hydroxylase activity that is dependent upon gonadotropic stimulation for expression.     

Mammary gland growth in the hypophysectomized pregnant rat (38522).

Sprague-Dawley-Rolfsmeyer rats were hypophysectomized on days 11, 12 or 15 of pregnancy and sacrificed on day 20 to determine the extent of mammary development, as assessed by determination of nucleic acid content. The DNA of six abdominal-inguinal glands in the hypophysectomized groups was not significantly different from that in the sham-operated pregnant or intact pregnant control groups. All groups maintaining pregnancy had significantly higher DNA contents in mammary glands than virgin control or hypophysectomized aborted groups. In order to determine the minimal numbers of placental-fetal units required to maintain pregnancy and mammary gland growth, fetuses and placentas were removed on day 12 of pregnancy in addition to the pituitary so that only one fetus and one placenta remained in the uterus of a group of 6 rats with other groups having 2, 3, 4, 5 remaining. Pregnancy was maintained with only one placental-fetal unit, but mammary gland proliferation was significantly lower than the control group on day 20 of pregnancy. Three to five conceptuses supported mammary proliferation during the latter half of pregnancy at a level not significantly different from intact or sham-operated control groups. Removal of placental units on day 12 in rats having pituitaries intact resulted in no mammary DNA change when 1-5 units remained. Removal of pitutaries on day 12 and placental-fetal units on day 14 also resulted in no change in mammary DNA with as little as two placentas (minus all fetuses),while only one placenta remaining resulted in a significantly lower mammary DNA than in groups wtih 2 or more placentas.