Expression of GH, TSH beta, LH beta and FSH beta genes during fetal pituitary development in the pig.

The development of the anterior pituitary gland involves the proliferation and differentiation of ectodermal cells in Rathke's pouch to generate distinct cell types, each of which produces its corresponding trophic hormone. Studying pituitary development will therefore reveal novel aspects of organogenesis. In the present study, we examined by in situ hybridization the expression of genes for anterior pituitary hormones during development of the fetal pig pituitary. We found that the beta-subunit gene of thyroid-stimulating hormone (TSH beta) was first expressed at E40, (E = day of embryonal/fetal life), growth hormone (GH) mRNA appeared between E40 and E50, and the gonadotrophin genes (LH beta and FSH beta) were expressed at E50. The transcripts for TSH beta, LH beta and FSH beta were abundantly expressed at about E80, while GH mRNA continued to be richly expressed until after birth. The GH gene was first expressed in the mantle layer of the anterior lobe, while the TSH beta and gonadotrophin (LH beta and FSH beta) mRNAs were found in the central and the basal regions of the anterior lobe, respectively. All of these mRNAs (GH, TSH beta, LH beta, and FSH beta) remained concentrated until the end of gestation in the area where they first appeared. The distinctive pattern of developmental expression of these hormone genes in the fetal pig anterior pituitary makes this tissue an excellent system in which to study tissue-specific gene activation and regulation.

[Somatomedins and their binding proteins are involved in wound healing after hypoxia of the central nervous system]. / Somatomedine und ihre Bindungsproteine sind an der Wundheilung nach Hypoxie im zentralen Nervensystem beteiligt.

Insulin-like growth factors (IGFs) are involved in cell growth and differentiation. In muscle tissue they regulate axonal in growth and maintain the connection. They also play a role in regeneration of the peripheral nerve system. We hypothesized that IGFs might also be important factors in the recovery of central nervous tissue after traumatic damage such as perinatal asphyxia. Our group developed a rat model to mimic the resulting damage and test the changes of expression of IGF-1, -2 and several of their binding proteins. We also examined the influence of exogenous IGF-1 and -2 after asphyxia in the same model. Rats underwent a unilateral ligation of the A. carotis followed by a 15 or 90 min inhalational hypoxia (8% O2). The treatment resulted in a mild or severe damage in the ligated hemisphere with either selective neuronal loss or complete infarction of the volume, respectively. The treatment induced expression of both IGFs and binding protein 2, 3 and 5. Binding protein 1 is not expressed and binding protein 4 is suppressed soon after hypoxia-ischemia. We conclude that both IGFs and several of their binding proteins are involved in response and wound healing after hypoxic brain damage. This was further tested in a second experiment. Rats were injected with IGF-1 or IGF-2 intra-ventricular soon after the hypoxic damage. IGF-1 treatment significantly reduced neuronal loss, IGF-2 had no effect. Behaviour tests, however, showed no difference between IGF-1 treated rats and controls. Our studies show interesting aspects for further investigation and a possible treatment of perinatal asphyxia and traumatic damage of nerve tissue.

Differential expression of insulin-like growth factor binding proteins (IGFBP) 4 and 5 mRNA in the rat brain after transient hypoxic-ischemic injury.

Recent studies suggest a role for the insulin-like growth factor (IGF) system in the repair of damaged tissue following hypoxic-ischemic injury in the infant rat brain. We have used a unilateral model of hypoxic-ischemic injury to assess the possible involvement of two IGF binding proteins (IGFBPs), IGFBP-4 and IGFBP-5, in the post-asphyxial response. Ligation of the right carotid artery of 21-day-old rats was followed by either 15 min or 60 min exposure to 8% oxygen to produce moderate and severe damage respectively. Using in situ hybridization, the distribution of IGFBP-4 and IGFBP-5 mRNA was determined in brains collected over 10 days following the insult. In the control brains (no damage), both IGFBPs were expressed in distinct regions. IGFBP-4 mRNA was detected in limited areas of the hippocampus and in several cortical layers, while IGFBP-5 mRNA was found primarily in the thalamus. In response to hypoxic-ischemic injury, IGFBP-4 mRNA expression was reduced in regions of neuronal loss, suggesting a neuronal origin for IGFBP-4. The expression of IGFBP-5 mRNA was not altered by the 15 min insult, but was heavily induced from 3 days following the 60 min insult, particularly in the subependymal layer and adjacent white matter on the ligated hemisphere. This suggests that IGFBP-5 may be involved in recovery from severe hypoxic-ischemic injury and may be important in the regeneration of oligodendrocytes.

Hypoxia and hypoxia/ischemia affect the expression of insulin-like growth factor binding protein 2 in the developing rat brain.

Brain halves were collected at various time points from 21-day-old Wistar rats exposed to either a short or prolonged period of inhalational hypoxia following unilateral carotid ligation post-insult. Neuronal loss was restricted to the side of the carotid ligation. Northern blot analysis was performed for IGFBP-2 mRNA. The prolonged hypoxia decreased the expression of IGFBP-2 five hours post insult, whereas the shorter insult level showed an (P < 0.05) increase above control. In both groups IGFBP-2 mRNA increased to peak 3-5 days post insult. After a decline at day 6, expression was again high 7-10 days after HI. In the short hypoxia group, where there was little neuronal loss, the expression of IGFBP-2 in both hemispheres followed the same pattern over time. However prolonged hypoxia induced higher IGFBP-2 expression in the ligated hemisphere where there was extensive neuronal loss. At day 5 post-insult 5/9 rats showed an additional, slightly smaller (1.4 kb vs 1.7 kb) second transcript. The different pattern of expression associated with different degrees of injury suggest that IGFBP-2 is involved in the post asphyxial response. Hypoxia itself leads to alterations in IGFBP-2 expression. Greater expression is associated with neuronal loss. These observations suggest that the IGF system contributes to neuronal rescue and/or brain repair processes.

Expression of insulin-like growth factor-binding protein 2 (IGF-BP 2) following transient hypoxia-ischemia in the infant rat brain.

Hypoxia-ischemia induced by unilateral carotid ligation followed by either 15 (moderate) or 90 (severe) min exposure to 8% oxygen was associated with induction of IGF-BP 2 mRNA expression. A specific rat IGF-BP 2 cDNA probe was used to determine the IGF-BP 2 mRNA distribution in brain sections using in situ hybridization. Untreated control rats and the non-ligated hemisphere in experimental rats expressed IGF-BP 2 mRNA in the choroid plexus, meninges and more weakly in the thalamus, hippocampus and cortical layer 5. Increased expression in experimental rats was limited to regions known to have neuronal damage. Three days after the moderate insult the signal was increased in the CA1/2 region of the hippocampus and thalamus of the ligated side. Three days after the severe insult IGF-BP 2 expression was found surrounding the infarcted regions while by 5 days after severe insult the whole infarcted volume showed induction. The results suggest a role for the IGFs in the post-asphyxial response. IGF-BP 2 may alter the bio-availability of IGF 1 or 2 or modulate their actions in the area of infarction, and thus promote cerebral repair and recovery.

Hypoxia-ischemia induces transforming growth factor beta 1 mRNA in the infant rat brain.

Transforming growth factor beta 1 (TGF beta 1) mRNA expression was examined after hypoxia-ischemia in rat brains using in situ hybridization. Twenty-one-day-old Wistar rats had unilateral ligation of the right carotid artery followed by either 15 or 90 min inhalational hypoxia. Fifteen min of hypoxia resulted in moderate damage with selective neuronal loss in cortical layer 3 and in the hippocampus of the ligated hemisphere. Seventy-two hours after hypoxia TGF beta 1 expression was markedly increased above control levels in those sites. Levels were normal after 120 h. Ninety min of hypoxia led to an infarction of the lateral cerebral cortex and hippocampus of the ligated hemisphere. One hour after hypoxia TGF beta 1 mRNA was expressed in the hippocampus of the damaged side. Seventy-two and 120 h after hypoxia, expressing cells were found throughout the cerebral cortex, piriform cortex, striatum, thalamus and hippocampus of the infarcted side. These data show that TGF beta 1 mRNA expression is induced after a hypoxic-ischemic insult in the brain. TGF beta 1 may be involved in post-asphyxial repair mechanisms.

A role for IGF-1 in the rescue of CNS neurons following hypoxic-ischemic injury.

Three days after unilateral hypoxic-ischemic injury in infant rats insulin-like growth factor 1 (IGF-1) production by astrocytes was enhanced in the injured region. This was associated with increased expression of mRNA for IGF binding protein-3 but not for binding protein-1. In adult rats a single lateral cerebroventricular injection of IGF-1 two hours following a similar injury markedly reduced neuronal loss. It is suggested that endogenous IGF-1 is neurotrophic and that centrally administered IGF-1 may have therapeutic potential for brain injury.