IGFBP-3在斑马鱼胚胎发育中的功能
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摘要
类胰岛素生长因子结合蛋白(IGFBP)-3 是血液中主要的 IGF 载体蛋白。 IGFBP-
3 延长了血液中的 IGFs 的半衰期并阻止了他们的潜在的低血糖效应。 IGFBP-3 也
表达在胎儿和成体阶段的外周组织中。 体外试验表明， IGFBP-3不仅能调节 IGF 的
生理作用， 还拥有不依赖于 IGF 的独立作用， 提示局部表达的 IGFBP-3 也具有旁
分泌和自分泌的功能。 然而， IGFBP-3 的在体功能尚不清楚。 本研究使用斑马鱼模
型探索了 IGFBP-3 的在胚胎发育中的功能。 IGFBP-3 mRNA 最早表达在迁移的头
神经嵴细胞， 随后表达在胚胎的咽弓和胸鳍原基。 IGFBP-3 mRNA 持续而强烈地
表达在发育中的内耳， 它也表达在眼睛的视杆和部分视网膜细胞， 松果腺以及脑垂
体前部、 中脑脑盖和下丘脑的基底中央等脑组织。 为了研究 IGFBP-3 在这些组织
中的功能， 我们使用 morpholino 修饰反义寡核苷酸(MOs) 阻断 IGFBP-3 基因产物
的表达。 IGFBP-3 knockdown 胚胎的咽弓骨骼形态发生滞后， 咽弓软骨分化明显
减少。 同时， IGFBP-3 knockdown 也引起内耳、 眼睛和胸鳍原基大小显著下降、 干
扰了内耳毛细胞和视神经的分化及半规管和松果腺的形成。 进一步， 我们在
knockdown胚胎中导入并表达 MO-抗性形式的 IGFBP-3“营救”了 MO-引起的缺
陷。 这些新的发现， 首次提供在体证据证明 IGFBP-3 在调节斑马鱼咽弓、 内耳的
生长发育、 视神经和运动神经的分化过程中起着重要作用， 并参与调节眼睛、 松果
腺、 胸鳍的发育过程。
Insulin-like growth factor binding protein (IGFBP)-3 is the major IGF carrier
protein in the bloodstream. The IGFBP-3 / IGF complex prolongs the half-life of IGFs in
circulation and prevents the potential hypoglycemic effect of IGFs. IGFBP-3 is also
expressed in many peripheral tissues in fetal and adult stages. In vitro studies using
cultured cells suggest that IGFBP-3 can inhibit / potentiate IGF actions, and even
possesses IGF-independent activities, suggesting that local IGFBP-3 may also act in a
paracrine / autocrine fashion. The in vivo function of IGFBP-3, however, is unknown. In
this study, we have explored the developmental role of IGFBP-3 using the zebrafish
model. RNA hybridization analyses revealed that IGFBP-3 mRNA expression began at
14 hour post fertilization (hpf) in the migrating cranial neural crest cells. It was later
transiently expressed in pharyngeal arches, and in pectoral fin buds. IGFBP-3 mRNA is
predominantly and persistently expressed in the developing otic vesicle. IGFBP-3 is also
expressed in optic stalk, retina, pineal gland, tegmentum in midbrain, anterior pituitary
and midline floor of hypothalamus. To determine the developmental role of IGFBP-3, we
ablated the IGFBP-3 gene product by injecting morpholino-modified antisense
oligonucleotides (MOs). The IGFBP-3 knocked-down morphants had delayed pharyngeal
skeleton morphogenesis and greatly reduced pharyngeal cartilage differentiation.
Knockdown of IGFBP-3 also significantly reduced sizes of otic vesicle, optic primordia,
pectoral fin bud, and disrupted the differentiations of hair cell and optic stalk, as well as
the formations of semicircular canal and pineal gland. Furthermore, re-introduction of a
MO-resistant form of IGFBP-3 “rescued” the MO-induced defects. This findings are the
first to provide in vivo evidences for IGFBP-3 playing an important role in regulating
development and growth of pharyngeal cartilage and otic vesicle, differentiations of optic
and motor neuron, as well as its involving in the development of eye, pineal gland and
pectoral fin bud in zebrafish.
3 延长了血液中的 IGFs 的半衰期并阻止了他们的潜在的低血糖效应。 IGFBP-3 也
表达在胎儿和成体阶段的外周组织中。 体外试验表明， IGFBP-3不仅能调节 IGF 的
生理作用， 还拥有不依赖于 IGF 的独立作用， 提示局部表达的 IGFBP-3 也具有旁
分泌和自分泌的功能。 然而， IGFBP-3 的在体功能尚不清楚。 本研究使用斑马鱼模
型探索了 IGFBP-3 的在胚胎发育中的功能。 IGFBP-3 mRNA 最早表达在迁移的头
神经嵴细胞， 随后表达在胚胎的咽弓和胸鳍原基。 IGFBP-3 mRNA 持续而强烈地
表达在发育中的内耳， 它也表达在眼睛的视杆和部分视网膜细胞， 松果腺以及脑垂
体前部、 中脑脑盖和下丘脑的基底中央等脑组织。 为了研究 IGFBP-3 在这些组织
中的功能， 我们使用 morpholino 修饰反义寡核苷酸(MOs) 阻断 IGFBP-3 基因产物
的表达。 IGFBP-3 knockdown 胚胎的咽弓骨骼形态发生滞后， 咽弓软骨分化明显
减少。 同时， IGFBP-3 knockdown 也引起内耳、 眼睛和胸鳍原基大小显著下降、 干
扰了内耳毛细胞和视神经的分化及半规管和松果腺的形成。 进一步， 我们在
knockdown胚胎中导入并表达 MO-抗性形式的 IGFBP-3“营救”了 MO-引起的缺
陷。 这些新的发现， 首次提供在体证据证明 IGFBP-3 在调节斑马鱼咽弓、 内耳的
生长发育、 视神经和运动神经的分化过程中起着重要作用， 并参与调节眼睛、 松果
腺、 胸鳍的发育过程。
Insulin-like growth factor binding protein (IGFBP)-3 is the major IGF carrier
protein in the bloodstream. The IGFBP-3 / IGF complex prolongs the half-life of IGFs in
circulation and prevents the potential hypoglycemic effect of IGFs. IGFBP-3 is also
expressed in many peripheral tissues in fetal and adult stages. In vitro studies using
cultured cells suggest that IGFBP-3 can inhibit / potentiate IGF actions, and even
possesses IGF-independent activities, suggesting that local IGFBP-3 may also act in a
paracrine / autocrine fashion. The in vivo function of IGFBP-3, however, is unknown. In
this study, we have explored the developmental role of IGFBP-3 using the zebrafish
model. RNA hybridization analyses revealed that IGFBP-3 mRNA expression began at
14 hour post fertilization (hpf) in the migrating cranial neural crest cells. It was later
transiently expressed in pharyngeal arches, and in pectoral fin buds. IGFBP-3 mRNA is
predominantly and persistently expressed in the developing otic vesicle. IGFBP-3 is also
expressed in optic stalk, retina, pineal gland, tegmentum in midbrain, anterior pituitary
and midline floor of hypothalamus. To determine the developmental role of IGFBP-3, we
ablated the IGFBP-3 gene product by injecting morpholino-modified antisense
oligonucleotides (MOs). The IGFBP-3 knocked-down morphants had delayed pharyngeal
skeleton morphogenesis and greatly reduced pharyngeal cartilage differentiation.
Knockdown of IGFBP-3 also significantly reduced sizes of otic vesicle, optic primordia,
pectoral fin bud, and disrupted the differentiations of hair cell and optic stalk, as well as
the formations of semicircular canal and pineal gland. Furthermore, re-introduction of a
MO-resistant form of IGFBP-3 “rescued” the MO-induced defects. This findings are the
first to provide in vivo evidences for IGFBP-3 playing an important role in regulating
development and growth of pharyngeal cartilage and otic vesicle, differentiations of optic
and motor neuron, as well as its involving in the development of eye, pineal gland and
pectoral fin bud in zebrafish.
引文
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