Computer-assisted quantitative immunohistochemistry of the
extracellular matrix in the dorsal and ventral spinal roots
with the use of a lucia software

J 2001

Computer-assisted quantitative immunohistochemistry of the extracellular matrix in the dorsal and ventral spinal roots with the use of a lucia software

DUBOVÝ, Petr; Ivana SVÍŽENSKÁ a Ilona KLUSÁKOVÁ

Základní údaje

Originální název

Computer-assisted quantitative immunohistochemistry of the extracellular matrix in the dorsal and ventral spinal roots with the use of a lucia software

Název česky

Komputerová analýza obrazu pro kvantitativní vyhodnocení imunohistochemie ECM

Autoři

DUBOVÝ, Petr; Ivana SVÍŽENSKÁ a Ilona KLUSÁKOVÁ

Vydání

The Histochemical Journal, Netherlands, Kluwer Academic Publishers, 2001, 0018-2214

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30000 3. Medical and Health Sciences

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 1.169

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14110/01:00005622

Organizační jednotka

Lékařská fakulta

Klíčová slova anglicky

quantitative immunohistochemistry; laminin; fibronectin

Štítky

fibronectin, laminin, quantitative immunohistochemistry

Změněno: 17. 6. 2009 11:01, prof. RNDr. Petr Dubový, CSc.

Anotace

ORIG CZ

V originále

The dorsal and ventral spinal roots contain different types of axons. The endoneurial extracellular matrix (ECM), produced by Schwann cells and fibroblasts under control of the axons, is important extrinsic factor of the nerve regeneration. A different content of the endoneurial ECM can be expected in the dorsal and ventral spinal roots in relation to their axon type differences, but no information is available to the present time. In the present paper we demonstrated a comparison of immunofluorescence intensity for chondroitin sulfate proteoglycan, fibronectin, tenascin and thrombospondin in the endoneurial ECM of the rat dorsal and ventral spinal roots. For this purpose the cryostat sections through the dorsal and ventral roots were cut and incubated simultaneously. We tested optimal incubation conditions with the primary and secondary antibodies to distinguish differences in immunostaining intensity of the endoneurial ECM for investigated molecules. The best results of distinct staining in the dorsal and ventral roots were obtained after incubation with the primary antibody for 4 hrs and after treatment with affinity purified secondary goat anti-mouse antibody conjugated with TRITC at room temperature (20-23oC) for 90 min. An intensity of the immunofluorescence staining was assessed by computer-assisted image analysis (Lucia-G v4.21) using interactive segmentation (HSI) to select the measured areas of digitized pictures. The connective tissue sheaths of both dorsal and ventral roots displayed a high intensity of immunofluorescence for all investigated molecules. The axoplasm of myelinated axons in both dorsal and ventral roots was slightly decorated by immunolabeling for chondroitin sulfate proteoglycan, tenascin and thrombospondin but not for fibronectin. The measurement of immunofluorescence brightness revealed that the endoneurial ECM of the dorsal roots is immunostained for investigated molecules in higher intensity than in the ventral roots. The results suggest quantitative differences of the endoneurial ECM content in the intact dorsal and ventral roots corresponding probably with presence of various types of axons. The different ECM contents of the intact spinal roots might be an initial stage related with different extrinsic conditions created alongside afferent and motor axons for promotion of their regeneration after the root injury.

Česky

Endoneuriální ECM je produkována Schwannovými buňkami a fibroblasty pod kontrolou axonů. Má rozdílné molekulární složení podél aferentních a eferentních nervových vláken.

Návaznosti

GA309/00/0407, projekt VaV

Název: Celulární a extracelulární prostředí pro regeneraci axonů při kořenových a distálních lezích periferních nervů

Investor: Grantová agentura ČR, Celulární a extracelulární prostředí pro regeneraci axonů při kořenových a distálních lezích periferních nervů

Citovat

DUBOVÝ, Petr; Ivana SVÍŽENSKÁ a Ilona KLUSÁKOVÁ. Computer-assisted quantitative immunohistochemistry of the extracellular matrix in the dorsal and ventral spinal roots with the use of a lucia software. The Histochemical Journal. Netherlands: Kluwer Academic Publishers, 2001, roč. 33, č. 8, s. 475-476. ISSN 0018-2214.

@article{401991,
   author = {Dubový, Petr and Svíženská, Ivana and Klusáková, Ilona},
   article_location = {Netherlands},
   article_number = {8},
   keywords = {quantitative immunohistochemistry; laminin; fibronectin},
   language = {eng},
   issn = {0018-2214},
   journal = {The Histochemical Journal},
   title = {Computer-assisted quantitative immunohistochemistry of the extracellular matrix in the dorsal and ventral spinal roots with the use of a lucia software},
   volume = {33},
   year = {2001}
}

TY  - JOUR
ID  - 401991
AU  - Dubový, Petr - Svíženská, Ivana - Klusáková, Ilona
PY  - 2001
TI  - Computer-assisted quantitative immunohistochemistry of the extracellular matrix in the dorsal and ventral spinal roots with the use of a lucia software
JF  - The Histochemical Journal
VL  - 33
IS  - 8
SP  - 475
EP  - 475
PB  - Kluwer Academic Publishers
SN  - 00182214
KW  - quantitative immunohistochemistry
KW  - laminin
KW  - fibronectin
N2  - The dorsal and ventral spinal roots contain different types of axons. The endoneurial extracellular matrix (ECM), produced by Schwann cells and fibroblasts under control of the axons, is important extrinsic factor of the nerve regeneration. A different content of the endoneurial ECM can be expected in the dorsal and ventral spinal roots in relation to their axon type differences, but no information is available to the present time. In the present paper we demonstrated a comparison of immunofluorescence intensity for chondroitin sulfate proteoglycan, fibronectin, tenascin and thrombospondin in the endoneurial ECM of the rat dorsal and ventral spinal roots. For this purpose the cryostat sections through the dorsal and ventral roots were cut and incubated simultaneously. We tested optimal incubation conditions with the primary and secondary antibodies to distinguish differences in immunostaining intensity of the endoneurial ECM for investigated molecules. The best results of distinct staining in the dorsal and ventral roots were obtained after incubation with the primary antibody for 4 hrs and after treatment with affinity purified secondary goat anti-mouse antibody conjugated with TRITC at room temperature (20-23oC) for 90 min. An intensity of the immunofluorescence staining was assessed by computer-assisted image analysis (Lucia-G v4.21) using interactive segmentation (HSI) to select the measured areas of digitized pictures. The connective tissue sheaths of both dorsal and ventral roots displayed a high intensity of immunofluorescence for all investigated molecules. The axoplasm of myelinated axons in both dorsal and ventral roots was slightly decorated by immunolabeling for chondroitin sulfate proteoglycan, tenascin and thrombospondin but not for fibronectin. The measurement of immunofluorescence brightness revealed that the endoneurial ECM of the dorsal roots is immunostained for investigated molecules in higher intensity than in the ventral roots. The results suggest quantitative differences of the endoneurial ECM content in the intact dorsal and ventral roots corresponding probably with presence of various types of axons. The different ECM contents of the intact spinal roots might be an initial stage related with different extrinsic conditions created alongside afferent and motor axons for promotion of their regeneration after the root injury.
ER  -

DUBOVÝ, Petr; Ivana SVÍŽENSKÁ a Ilona KLUSÁKOVÁ. Computer-assisted quantitative immunohistochemistry of the extracellular matrix in the dorsal and ventral spinal roots with the use of a lucia software. \textit{The Histochemical Journal}. Netherlands: Kluwer Academic Publishers, 2001, roč.~33, č.~8, s.~475-476. ISSN~0018-2214.

Přehled o publikaci