Monocytes may promote myofibroblast accumulation and apoptosis in Alport renal fibrosis.

BACKGROUND: In interstitial fibrosis, monocytes and myofibroblasts have been directly implicated in scarring, apoptosis, and tissue necrosis. While much has been done to explore the role of these cell types individually in fibrosis, the interactive dependency of monocytes and myofibroblasts has been only marginally explored. METHODS: Alport mice were treated or not with a soluble receptor inhibitor for transforming growth factor-beta 1 (TGF-beta 1), which was previously shown to inhibit the accumulation of myofibroblasts, but not monocytes, in the tubulointerstitium. Kidneys were examined for fibrosis using several matrix markers, TGF-beta 1 mRNA expression by in situ hybridization, apoptosis using the terminal deoxynucleotidyl transferase-mediated uridine triphosphate nick end labeling (TUNEL) assay, expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPS) by dual immunofluorescence microscopy, MMP activity by gelatin and in situ zymography, MMP mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR), and basement membrane degradation by dual immunofluorescence confocal microscopy and electron microscopy. RESULTS: Treated mice showed a markedly reduced accumulation of matrix proteins. Tissue monocytes express TGF-beta 1 mRNA, and TGF-beta 1 is required for myofibroblast accumulation. The number of apoptotic cells was not influenced by TGF-beta 1 inhibition. Monocytes express MMP-2, MMP-9, TIMP-2, and TIMP-3. MMP activity and mRNA expression is equally up regulated in treated and untreated Alport mice. Tubular basement membranes (TBM) around clusters of monocytes are notably degraded. TGF-beta 1 inhibition does not extend the life of Alport mice. CONCLUSION: These studies demonstrate that monocytes may influence myofibroblast accumulation via TGF-beta1, and that monocytes, and not myofibroblasts, are associated with tubular atrophy in Alport mice. Elevated MMP expression and activity is associated with TBM destruction near monocytes clusters, suggesting an anoikis mechanism may contribute to apoptosis in this model.

Functional integrity of ErbB-4/-2 tyrosine kinase receptor complex in the hypothalamus is required for maintaining normal reproduction in young adult female rats.

ErbB-1 tyrosine kinase receptors are necessary for maintaining female reproduction by modulating the release of LH-releasing hormone (LHRH). Changes in ErbB-1 signaling capacity in aging rats are linked to compromised reproduction. The interactive and synergistic nature of different members of ErbB receptors in mediating signal transduction exists in many cellular systems. Particularly, the interactions among ErbB-1 and ErbB-2 or ErbB-4 and ErbB-2 are known to be involved in the stimulation of LHRH secretion during sexual maturation. Thus, ErbB-4/-2 receptors may also play a role in maintaining reproduction during adulthood, and consequently, alteration in ErbB-4/-2 signaling capacity may contribute to compromised reproductive competence during aging. By in situ hybridization histochemistry, ErbB-4/-2 mRNAs were detected in the preoptic area (POA) and arcuate nucleus, which are important areas involved in the control of LHRH neuronal activity. RT-PCR analyses showed that levels of ErbB-4/-2 mRNA increased to a maximal value in the POA of young adult animals before the LH surge. However, no such increase was found in middle-aged female rats. The timing of the decrease in ErbB-4 mRNA in the median eminence-arcuate nucleus of middle-aged rats was delayed compared with that in young adult animals. Disruption of functional ErbB-4/-2 receptor complex by blocking ErbB-2 receptor synthesis in the hypothalamus via an infusion of ErbB-2 antisense oligodeoxynucleotide resulted in an estrous acyclicity in young adult rats. These results indicate that changes in ErbB-4/-2 gene expression and functional integrity of this ErbB-4/-2 receptor complex in the hypothalamus of middle-aged female animals may lead to an altered preovulatory LH release. Thus, the ErbB-4/-2 receptor complex is a physiological component necessary for maintaining female reproduction.

Altered gene activity of epidermal growth factor receptor (ErbB-1) in the hypothalamus of aging female rat is linked to abnormal estrous cycles.

Activation of the ErbB-1 receptor is necessary for initiating mammalian female puberty by stimulating the release of LH-releasing hormone. It remains unclear whether ErbB-1 is also required in governing reproduction during adulthood and whether altered ErbB-1 signaling is linked to changes in gonadotropin secretion in aging females. The present study examined these issues. RT-PCR was employed to determine changes in ErbB-1 mRNA levels during proestrus in both young adult (YA) and middle-aged (MA) female rats. Before the LH surge, expression levels in the preoptic area of YA rats increased to a maximal value. No such increase in ErbB-1 mRNA was found in MA rats. This difference was confirmed by the analysis of in situ hybridization histochemistry, where a stronger mRNA signal was observed in the preoptic area of YA rats compared with MA females. ErbB-1 protein levels measured by Western blot reflected this difference. A peak level of ErbB-1 mRNA in the median eminence-arcuate nucleus was detected at 0800 h in YA rats, but it was delayed in MA animals. There were intense ErbB-1 mRNA-positive cells in the arcuate nucleus. Pharmacological blockade of ErbB-1 receptor-mediated signal transduction resulted in the disruption of estrous cyclicity in YA rats. These results indicate that ErbB-1 receptors are necessary for maintaining normal estrous cycles. Consequently, age-related alterations in hypothalamic ErbB-1 gene activity may contribute to a delayed preovulatory LH secretion in aging females. Thus, the ErbB-1 signaling system plays an important role in the control of female reproduction during adulthood.