NGF-like trophic support from peripheral nerve for grafted rhesus adrenal chromaffin cells

Abstract

Autopsy results on patients and corresponding studies in nonhuman primates have revealed that autografts of adrenal medulla into the striatum, used as a treatment for Parkinson's disease, do not survive well. Because adrenal chromaffin cell viability may be limited by the low levels of available nerve growth factor (NGF) in the striatum, the present study was conducted to determine if transected peripheral nerve segments could provide sufficient levels of NGF to enhance chromaffin cell survival in vitro and in vivo. Aged female rhesus monkeys, rendered hemiparkinsonian by the drug MPTP (n-methyl-4-phenyl-1,2,3,6 tetrahydropyridine), received autografts into the striatum using a stereotactic approach, of either sural nerve or adrenal medulla, or cografts of adrenal medulla and sural nerve (three animals in each group). Cell cultures were established from tissue not used in the grafts. Adrenal chromaffin cells either cocultured with sural nerve segments or exposed to exogenous NGF differentiated into a neuronal phenotype. Chromaffin cell survival, when cografted with sural nerve into the striatum, was enhanced four- to eightfold from between 8000 and 18,000 surviving cells in grafts of adrenal tissue only up to 67,000 surviving chromaffin cells in cografts. In grafts of adrenal tissue only, the implant site consisted of an inflammatory focus. Surviving chromaffin cells, which could be identified by both chromogranin A and tyrosine hydroxylase staining, retained their endocrine phenotype. Cografted chromaffin cells exhibited multipolar neuritic processes and numerous chromaffin granules, and were also immunoreactive for tyrosine hydroxylase and chromogranin A. Blood vessels within the graft were fenestrated, indicating that the blood-brain barrier was not intact. Additionally, cografted chromaffin cells were observed in a postsynaptic relationship with axon terminals from an undetermined but presumably a host origin.

Robust survival of isolated bovine adrenal chromaffin cells following intrastriatal transplantation: a novel hypothesis of adrenal graft viability

“…The experimental basis for this hypothesis principally stems from the observation that NGF supports the viability and phenotypic differentiation of adrenal chromaffin cells in vitro (Unsicker et al, 1978; see also Notter et al, 1989;Kordower et al, 1990a,b) and following intracerebra1 grafting (see reviews by Kordower et al, 1990bKordower et al, , 1992. Regarding neural grafting, the seminal study of Stromberg et al (1985) demonstrated that large numbers of adrenal chromaffin cells survive for up to 1 year within the striatum, become mor- phologically differentiated, and potentiate functional recovery when the implant receives injections of P-NGF.…”

“…We and others have demonstrated that cografting adrenal chromaffin cells with biological sources of NGF also enhances the viability of grafted chromaffin cells. In this regard, chromaffin cells cografted with growth factor-producing C6 gliomas (Bing et al, 1990), astrocytes or fibroblasts genetically engineered to produce NGF (Cunningham et al, 199 1;Chalmers et al, 1992), or growth factorproducing transected peripheral nerve (Date et al, 1990a,b;Kordower et al, 1990b;Doering, 199 1) potentiate the viability, and in some cases functionality, of adrenal medullary grafts. In addition, the improved survival of adrenal medullary grafts placed into brain regions that synthesize high levels of endogenous NGF such as the hippocampus (Jousselin-Hosaja, 1988a,b) and cerebral cortex (Morihisa et al, 1984) further supports the notion that low striatal levels of NGF are responsible for the poor survival of intrastriatal implants.…”

“…Sections were processed for the light microscopic visualization of tyrosine hydroxylase (TH), dopamine p-hydroxylase (DfiH), phenylethanolamine n-methyl transferase (PNMT), and chromogranin A according to previously published procedures (Fiandata et al, 1988;Kordower et al, 1988Kordower et al, , 1990b. Briefly, endogenous peroxidase activity was eliminated with a 20 min incubation in 0.1 M sodium periodate.…”

Enhanced survival and neuronal differentiation of adrenal chromaffin cells cografted into the striatum with NGF-producing fibroblasts

“…Survival of chromaffin cells transplanted into the brain parenchyma is, however, generally poor (Freed et al, 1986;Bohn et al, 1987;Hansen et al, 1988;Hurtig et al, 1989;Jankovic et al, 1989;Peterson et al, 1989;Hirsch et al, 1990;Kordower et al, 199 1). To increase the survival of grafted chromaffin cells, supplementation with NGF has been successfully employed (Stromberg et al, 1985;Date et al, 1990;Kordower et al, 1990;Cunningham et al, 1991). In addition to increasing chromaffin cell survival, NGF also induces the chromaffin cells to develop neurites (Stromberg et al, 1985;Date et al, 1990;Kordower et al, 1990;Cunningham et al, 1991).…”

Promoter-activated expression of nerve growth factor for treatment of neurodegenerative diseases

“…The potential utility of established cell lines as donor tissue for neural implants has been demonstrated in several species, including non-human primates; 19 IMR-32 neuroblastoma cells, rendered amitotic and differentiated in vitro, survived for over a year following hippocampal implantation in fimbria-fornix lesioned monkeys. Furthermore, the addition of exogenous trophic factors such as NGF or basic fibroblast growth factor (bFGF) enhances survival and function of grafted adrenal chromaffin cells, 20,21 and protects septo-hippocampal cholinergic neurons from lesion-induced degeneration 22,23 and from glucose deprivation-induced neurofibrillar tangle expression.…”