On the actions of neurotrophic factors on the chromaffin cells of the adrenal medulla
Author: Förander, Petter
Date: 2000-05-05
Location: Hörsalen, Enheten för hygien, Nobels väg 11
Time: 9.00
Department: Institutionen för neurovetenskap / Department of Neuroscience
Abstract
Chromaffin cells producing adrenaline or noradrenaline are the predominant cell type in the adrenal medulla, but ganglion cells and Schwann cells are also present. All cells of the adrenal medulla are derived from the neural crest, which is a transient neuroectoderm-derived embryonic structure. During the migration of the pluripotent neural crest cells they differentiate into their mature phenotype. The differentiation is regulated by signal molecules, such as neurotrophic factors and steroid hormones. The pluripotency of the neural crest cells is to some extent maintained in the mature chromaffin cells. In this thesis chromaffin cells were used to study the effects of nerve growth factor (NGF) and other neurotrophic factors on the transformation of adult chromaffin cells into neuronal-like phenotypes, as well as the expression and regulation of neurotrofic factors and their receptors. Furthermore, striatal neuronal responses to intraparenchymal infusion of NGF was studied. In situ hybridization, immunohistochemistry, neurite outgrowth assay, TGFß and NGF assays, and HPLC were used to study the chromaffin cells in situ and the effects of neurotrophic factors on intraocular transplanted and primary cultured adult chromaffin cells.
Dose-dependent reinnervation of host irides in response to different concentrations of NGF was shown. Nerve fiber formation was saturated when transplants were treated with NGF (100 [my]g/ml), and the half-maximal effect was achieved when host eyes were injected with 30 [my]g/ml NGF. As a result of the first study a baseline of neurite formation in response to NGF had been documented, and therefore other neurotrophic factors were tested using the same experimental model. The results revealed that ciliary neurotrophic factor (CNTF), basic fibroblast growth factor (bFGF), neurotrophin 4 (NT-4), glial cell line-derived neurotrophic factor (GDNF), at concentrations of 100200 [my]g/ml, stimulated nerve fiber formation from chromaffin grafts. Neurotrophin 3 (NT-3) was the only neurotrophic factor tested that had no effect on neurite outgrowth.
In the next series of experiment the relationship between NGF and transforming growth factor-ß (TGFß) was studied on the chromaffin cells. No synergistic effect was found on neurite formation after co-administration of TGFß and NGF in the chromaffin grafts. Instead grafts treated with NGF and antibodies against TGFß resulted in enlarged area of reinnervation. TGFß and NGF had thus antagonistic effects on neurite formation from the chromaffin cells. Using in situ hybridization CNTF receptor alpha (CNTFR[alpha]) mRNA was detected in P1, and adult adrenal medullary cells. Increased level of CNTFR[alpha] mRNA was seen after NGF treatment. Furthermore, GDNF family receptor alpha I (GFR[alpha]1) mRNA was present in the ganglion cells of the adrenal medulla, when adult adrenal medullary tissue was explanted, many chromaffin cells became GFR[alpha]1 mRNA-positive. Factors within cell culture medium were important for the expression of GFR[alpha]1 in the cultured chromaffin cells. GDNF induced neurite formation from intraocular chromaffin grafts, and this neurite-promoting capacity of GDNF, was confirmed in primary cultures of adult chromaffin cells.
Intrastriatal transplants of chromaffin tissue as a source of catecholamines, have been proposed as an alternative treatment strategy in Parkinsion's disease. In clinical trials intrastriatal chromaffin transplant have been stimulated by co-administration of NGF. A transient reduction of some typical parkinsonian symptoms was reported. In the last article of this thesis NGF was intraparenchymally infused into the normal rat striatum, to evaluate the role of NGF on the striatal neurons. NGF diffused from the infusion site to the overlying cortex and to the hippocampus. The results revealed that increased levels of mRNA corresponding to trkA, p75, muscarin receptor 2, ChAT and increased levels of acetylcholine esterase in the NGF infused striatum compared to contralateral side were found. Thus, NGF stimulates the cholinergic system, which is believed to be excitatory in the striatum. Surprisingly, a decreased spontaneous firing rate was found in the NGF infused animals, although the cholinergic markers were upregulated. This downregulation of spontaneous activity in the striatum could explain some of the beneficial effects, seen in patients suffering from Parkinson's disease, when NGF was co-administrated to the intra striatal chromaffin graft.
Dose-dependent reinnervation of host irides in response to different concentrations of NGF was shown. Nerve fiber formation was saturated when transplants were treated with NGF (100 [my]g/ml), and the half-maximal effect was achieved when host eyes were injected with 30 [my]g/ml NGF. As a result of the first study a baseline of neurite formation in response to NGF had been documented, and therefore other neurotrophic factors were tested using the same experimental model. The results revealed that ciliary neurotrophic factor (CNTF), basic fibroblast growth factor (bFGF), neurotrophin 4 (NT-4), glial cell line-derived neurotrophic factor (GDNF), at concentrations of 100200 [my]g/ml, stimulated nerve fiber formation from chromaffin grafts. Neurotrophin 3 (NT-3) was the only neurotrophic factor tested that had no effect on neurite outgrowth.
In the next series of experiment the relationship between NGF and transforming growth factor-ß (TGFß) was studied on the chromaffin cells. No synergistic effect was found on neurite formation after co-administration of TGFß and NGF in the chromaffin grafts. Instead grafts treated with NGF and antibodies against TGFß resulted in enlarged area of reinnervation. TGFß and NGF had thus antagonistic effects on neurite formation from the chromaffin cells. Using in situ hybridization CNTF receptor alpha (CNTFR[alpha]) mRNA was detected in P1, and adult adrenal medullary cells. Increased level of CNTFR[alpha] mRNA was seen after NGF treatment. Furthermore, GDNF family receptor alpha I (GFR[alpha]1) mRNA was present in the ganglion cells of the adrenal medulla, when adult adrenal medullary tissue was explanted, many chromaffin cells became GFR[alpha]1 mRNA-positive. Factors within cell culture medium were important for the expression of GFR[alpha]1 in the cultured chromaffin cells. GDNF induced neurite formation from intraocular chromaffin grafts, and this neurite-promoting capacity of GDNF, was confirmed in primary cultures of adult chromaffin cells.
Intrastriatal transplants of chromaffin tissue as a source of catecholamines, have been proposed as an alternative treatment strategy in Parkinsion's disease. In clinical trials intrastriatal chromaffin transplant have been stimulated by co-administration of NGF. A transient reduction of some typical parkinsonian symptoms was reported. In the last article of this thesis NGF was intraparenchymally infused into the normal rat striatum, to evaluate the role of NGF on the striatal neurons. NGF diffused from the infusion site to the overlying cortex and to the hippocampus. The results revealed that increased levels of mRNA corresponding to trkA, p75, muscarin receptor 2, ChAT and increased levels of acetylcholine esterase in the NGF infused striatum compared to contralateral side were found. Thus, NGF stimulates the cholinergic system, which is believed to be excitatory in the striatum. Surprisingly, a decreased spontaneous firing rate was found in the NGF infused animals, although the cholinergic markers were upregulated. This downregulation of spontaneous activity in the striatum could explain some of the beneficial effects, seen in patients suffering from Parkinson's disease, when NGF was co-administrated to the intra striatal chromaffin graft.
List of papers:
I. Förander P, Björklund L, Strömberg I (1994). Dose-dependent effects of recombinant human NGF on grafted adult adrenal medullary tissue. Exp Neurol. 126(2): 168-77.
Pubmed
II. Förander P, Hoffer B, Strömberg I (1998). Nerve fiber formation and catecholamine content in adult rat adrenal medullary transplants after treatment with NGF, NT-3, NT-4/5, bFGF, CNTF, and GDNF. Cell Tissue Res. 292(3): 503-512.
Pubmed
III. Förander P, Krieglstein K, Söderström S, Strömberg I (2000). Mutual induction of TGFbeta1 and NGF after treatment with NGF or TGFbeta1 in grafted chromaffin cells of the adrenal medulla. Exp Neurol. 164(2):303-13.
Pubmed
IV. Förander P, Brené S, Strömberg I (2000). Expression and regulation of CNTF-receptor-alpha in the in situ and in oculo grafted rat adrenal medulla. Neuroreport. 11: 1-5.
Pubmed
V. Förander P, Broberger C, Johansson M, Strömberg I (2000). GNDF induces neuronal transformation in primary cultures of adrenal chromaffin cells. [Submitted]
VI. Förander P, Söderström S, Humpel C, Strömberg I (1996). Chronic infusion of nerve growth factor into rat striatum increases cholinergic markers and inhibits striatal neuronal discharge rate. Eur J Neurosci. 8(9): 1822-32.
Pubmed
I. Förander P, Björklund L, Strömberg I (1994). Dose-dependent effects of recombinant human NGF on grafted adult adrenal medullary tissue. Exp Neurol. 126(2): 168-77.
Pubmed
II. Förander P, Hoffer B, Strömberg I (1998). Nerve fiber formation and catecholamine content in adult rat adrenal medullary transplants after treatment with NGF, NT-3, NT-4/5, bFGF, CNTF, and GDNF. Cell Tissue Res. 292(3): 503-512.
Pubmed
III. Förander P, Krieglstein K, Söderström S, Strömberg I (2000). Mutual induction of TGFbeta1 and NGF after treatment with NGF or TGFbeta1 in grafted chromaffin cells of the adrenal medulla. Exp Neurol. 164(2):303-13.
Pubmed
IV. Förander P, Brené S, Strömberg I (2000). Expression and regulation of CNTF-receptor-alpha in the in situ and in oculo grafted rat adrenal medulla. Neuroreport. 11: 1-5.
Pubmed
V. Förander P, Broberger C, Johansson M, Strömberg I (2000). GNDF induces neuronal transformation in primary cultures of adrenal chromaffin cells. [Submitted]
VI. Förander P, Söderström S, Humpel C, Strömberg I (1996). Chronic infusion of nerve growth factor into rat striatum increases cholinergic markers and inhibits striatal neuronal discharge rate. Eur J Neurosci. 8(9): 1822-32.
Pubmed
Issue date: 2000-04-14
Publication year: 2000
ISBN: 91-628-4145-9
Statistics
Total Visits
Views | |
---|---|
On ...(legacy) | 505 |
On ... | 234 |
Total Visits Per Month
October 2023 | November 2023 | December 2023 | January 2024 | February 2024 | March 2024 | April 2024 | |
---|---|---|---|---|---|---|---|
On ... | 3 | 7 | 0 | 84 | 0 | 0 | 1 |
Top country views
Views | |
---|---|
United States | 164 |
China | 155 |
Sweden | 62 |
Germany | 44 |
South Korea | 38 |
France | 9 |
Poland | 8 |
Denmark | 6 |
Finland | 6 |
Hong Kong | 5 |
Top cities views
Views | |
---|---|
Dongguan | 84 |
Sunnyvale | 32 |
Seoul | 30 |
Beijing | 22 |
Kiez | 19 |
Shenzhen | 15 |
Stockholm | 11 |
Wilmington | 9 |
Kraków | 8 |
Tianjin | 7 |