• Applied Microscopy
• /
• 제26권2호
• /
• pp.137-155
• /
• 1996

The development of the ganglia of the trachea was studied by electron microscopy in human fetuses ranging from 40 mm to 260 mm crown rump length. At 40 mm fetus, the tracheal ganglia was observed in the submucosa of the trachea. The primitive ganglia consisted of neuroblasts, undifferentiated cells, and unmyelinated nerve fibers. At 50 mm fetus, the neuroblast and their processes in the tracheal ganglia ware ensheathed by the bodies or processes of satellite cells. The cytoplasm of the neuroblast contained rough endoplasmic reticulum, mitochondria, Golgi complex, and ribosomes. At 70 mm fetus, cholinergic and adrenergic axon terminals were observed. Cholinergic axon terminals with agranular vesicles were abundant in the tracheal ganglia with increasing age. During next prenatal stage from 100 mm fetus, the ganglion cells and its processes were completely covered by a thin processes of the satellite cells. Unmyelinated nerve fibers were also completely ensheathed by processes of Schwann cell. Synaptic contacts between the cholinergic axon and dendrite of ganglion cells and a few dendrodendritic synapses were first observed at 100 mm fetus. The granule-containing cells were first identified in the tracheal ganglia at 200 mm fetus. These findings indicate that tracheal ganglia of human fetus resembles other parasympathetic and sympathetic ganglia, but not the enteric ganglia.

• Anatomy and Cell Biology
• /
• 제51권4호
• /
• pp.266-273
• /
• 2018

The ganglion cardiacum or juxtaductal body is situated along the left recurrent laryngeal nerve in the aortic window and is an extremely large component of the cardiac nerve plexus. This study was performed to describe the morphologies of the ganglion cardiacum or juxtaductal body in human fetuses and to compare characteristics with intracardiac ganglion. Ganglia were immunostained in specimens from five fetuses of gestational age 12-16 weeks and seven fetuses of gestational age 28-34 weeks. Many ganglion cells in the ganglia were positive for tyrosine hydroxylase (TH; sympathetic nerve marker) and chromogranin A, while a few neurons were positive for neuronal nitric oxide synthase (NOS; parasympathetic nerve marker) or calretinin. Another ganglion at the base of the ascending aorta carried almost the same neuronal populations, whereas a ganglion along the left common cardinal vein contained neurons positive for chromogranin A and NOS but no or few TH-positive neurons, suggesting a site-dependent difference in composite neurons. Mixtures of sympathetic and parasympathetic neurons within a single ganglion are consistent with the morphology of the cranial base and pelvic ganglia. Most of the intracardiac neurons are likely to have a non-adrenergic non-cholinergic phenotype, whereas fewer neurons have a dual cholinergic/noradrenergic phenotype. However, there was no evidence showing that chromogranin A- and/or calretinin-positive cardiac neurons corresponded to these specific phenotypes. The present study suggested that the ganglion cardiacum was composed of a mixture of sympathetic and parasympathetic neurons, which were characterized the site-dependent differences in and near the heart.

• The Korean Journal of Physiology and Pharmacology
• /
• 제6권2호
• /
• pp.63-70
• /
• 2002

Cholinergic modulation of GABAergic spontaneous miniature inhibitory postsynaptic currents (mIPSCs) by the activation of muscarine receptors was investigated in mechanically dissociated rat nucleus basalis of the Meynert neurons using the conventional whole-cell patch recording configuration. Muscarine $(10{\mu}M)$ reversibly and concentration-dependently decreased mIPSC frequency without affecting the current amplitude distribution. Muscarine action on GABAergic mIPSCs was completely blocked by $1{\mu}M$ methoctramine, a selective $M_2$ receptor antagonist, but not by $1{\mu}M$ pirenzepine, a selective $M_1$ receptor antagonist. NEM $(10{\mu}M),$ a G-protein uncoupler, attenuated the inhibitory action of muscarine on GABAergic mIPSC frequency. Muscarine still could decrease GABAergic mIPSC frequency even in the $Ca^{2+}-free$ external solution. However, the inhibitory action of muscarine on GABAergic mIPSCs was completely occluded in the presence of forskolin. The results suggest that muscarine acts presynaptically and reduces the probability of spontaneous GABA release, and that such muscarine-induced inhibitory action seems to be mediated by G-protein-coupled $M_2$ receptors, via the reduction of cAMP production. Accordingly, $M_2$ receptor-mediated disinhibition of nBM neurons might play one of important roles in the regulation of cholinergic outputs from nBM neurons as well as the excitability of nBM neurons themselves.

• The Korean Journal of Pain
• /
• 제11권1호
• /
• pp.7-22
• /
• 1998

The development of the superior cervical ganglion was studied by electron microscopic method in human fetuses ranging from 40 mm to 260 mm of crown-rump length(10 to 30 weeks of gestational age). At 40 mm fetus, the superior cervical ganglion was composed of clusters of undifferentiated cell, primitive neuroblast, primitive supporting cell, and unmyelinated fibers. At 70 mm fetus, the neuroblasts and their processes were ensheated by the bodies or processes of satellite cells. The cytoplasm of the neuroblast contained rough endoplasmic reticulum, mitochondria, Golgi complex, Nissl bodies and dense-cored vesicles. As the neuroblasts grew and differentiated dense-cored vesicles moved away from perikaryal cytoplasm into developing processes. Synaptic contacts between the cholinergic axon and dendrites of postganglionic neuron and a few axosomatic synapses were first observed at 70 mm fetus. At 90 mm fetus the superior cervical ganglion consisted of neuroblasts, satellite cells, granule-containing cells, and unmyelinated nerve fibers. The ganglion cells increased somewhat in numbers and size by 150 mm fetus. Further differentiation resulted in the formation of young ganglion cells, whose cytoplasm was densely filled with cell organelles. During next prenatal stage up to 260 mm fetus, the cytoplasm of the ganglion cells contained except for large pigment granules, all intracytoplasmic structures which were also found in mature superior cervical ganglion. A great number of synaptic contact zones between the cholinergic preganglionic axon and the dendrites of the postganglionic neuron were observed and a few axosomatic synapses were also observed. Two morphological types of the granule-containing cells in the superior cervical ganglion were first identified at 90 mm fetus. Type I granule-containing cell occurred in solitary, whereas type II tended to appeared in clusters near the blood capillaries. Synaptic contacts were first found on the solitary granule-containing cell at 150 mm fetus. Synaptic contacts between the soma of type I granule-containing cells and preganglionic axon termials were observed. In addition, synaptic junctions between the processes of the granule-containing cells and dendrites of postganglionic neuron were also observed from 150 mm fetus onward. In conclusion, superior cervical ganglion cells and granule-containing cells arise from a common undifferentiated cell precursor of neural crest. The granule-containg cells exhibit a local modulatory feedback system in the superior cervical ganglion and may serve as interneurons between the preganglionic and postganglionic cells.

• 한국동물학회지
• /
• 제38권2호
• /
• pp.248-268
• /
• 1995

출생 후 0일, 7일, 14일, 21일 그리고 성체의 흰쥐 전뇌 기저부의 내측중격핵과 수직 및 수평대각 Broca대에서 choline acetyltransferase(ChAT)에 면역반응을 보이는 신경조직과 세포의 분화를 면역조직화학적 및 전자현미경적 방법을 이용하여 조사하였다. 출생 후 초기와 성체에서 신경세포의 세포질과 수상돌기에서 고루 ChAT 면역반응이 확인되었다. 뇌 기저부의 ChAT 면역반응 신경세포들은 발생에 따른 뇌 크기의 증대와 뇌 조직의 분화에 따라 점차 수적 증가를 보였다. 이 ChAT 면역반응 신경세포들은 세포의 모양과 세포제의 장 · 단축의 비에 따라 6가지 형 즉 1) 월형 2) 난형, 3) 세장형, 4) 방추형. 5) 삼각형, 6) 다각형으로 분류되었다 전뇌 기저 핵에서 원형과 난형 신경세포들의 출현율은 출생 후 0일에서 높았으나 성체로 되면서 감소된 반면, 세장형. 방추형. 삼각형 그리고 다각형 신경세포들의 출현율은 출생 후 0일에서는 낮았으나 성체로 되면서 증가하였다. 모든 핵들에서 ChAT 면역반응 신경세포체의 부피는 출생 후 0일에 996-1,252 Um3로 제일 작았으며, 내측중격핵과 수직대각 Broca대 그리고 수평대각 Broca대에서는 출생 후 21일에 각각 5,061, 5.701, 5,820 um3로 최대치를 보였다. 그후 성체로 되면서 모든 핵에서1,897-2,704 roms로 다시 감소하였다. 전자현미경적 관찰에서 출생 후 21일된 흰쥐 수평대각 Broca대에서 ChAT 면역반응은 핵의 핵질 일부와 핵막 그리고 미토콘드리아와 조면소포체에서 관찰되었다. 이 결과들로 미루어 출생 후 초기 발생단계에서 흰쥐 전뇌 기저부의 내측중격핵과 수직 및 수평대각 Broca대에서 ChAT 면역반응 신경세포들은 축삭과 수상돌기의 형성에 따라 세포의 형과 그 출현율 및 세포제의 크기에서 현저한 변화가 이루어지는 것으로 생각된다.

• Applied Microscopy
• /
• 제31권4호
• /
• pp.355-365
• /
• 2001

흰쥐 전뇌 기저부에서 콜린성 신경세포는 내측중격핵, 수직 및 수평 대각 Broca대, 거대세포 시교차앞핵, 복부담창구, Meynert기저핵 그리고 신선조체 등에 분포하는 것으로 알려져 있다. 저자 등은 흰쥐 전뇌 기저부 Meynert 기저핵에서 ChAT항체를 이용한 면역조직 및 세포화학적 방법으로 신경세포들의 발생, 분포, 형태, 형별 출현율 및 세포체의 부피, 조직학적 특성 등을 확인하였다. 또한 전자현미경을 이용하여 ChAT항체에 면역 반응을 하는 신경세포의 미세구조적 특징을 관찰하여 출생 후 발생단계에 따른 Meynert 기저핵에서 ChAT항체 면역반응 신경세포들의 분화를 관찰하고 다음과 같은 결과를 얻었다. ChAT항체에 반응하는 신경세포의 분포는 조직마다 부분적으로 상이하지만, 대부분 담창구가 내포와 접하는 부위, 또는 담창구와 무명질의 연접지역에 군집을 이루거나 혹은 단독으로 관찰되었으며 ,무명질에는 시교차앞핵의 면역반응 세포와 혼재하는 것으로 확인되었다. ChAT면역반응 신경세포들은 세포의 모양과 세포체의 장 단축의 비에 따라 6가지형으로 분류되었다. 원형의 신경세포 출현율은 출생 후 7일에서 가장 높았고, 성체로 되면서 점차적으로 감소하였으며, 난형 신경세포는 출생 후 14일에 가장 높게 나타났고, 원형의 신경세포와 같이 성체로 되면서 점차 감소하였다. 또한, 세장형 신경세포의 출현 율은 출생 후 21일에 가장 높게 나타났다. 삼각형, 방추형 그리고 다각형 신경세포는 성체에서 출현율이 높았다. ChAT면역반응 신경세포체의 부피는 출생 후 7일에 $1,268{\mu}m^3$로 제일 작았으나, 출생 후 21일에서는 $4,453{\mu}m$로 최대치를 보였다. 그 후 성체로 되면서 점차 감소되었다. 전자현미경 관찰에서 성체의 ChAT항체 반응 신경세포의 핵외막, rER의 막, 자유리보즘 그리고 polysorne등이 관찰되었으며, 대칭 및 비대칭 신경연접이 관찰되었다. 이상의 결과들로 미루어 흰쥐 전뇌 기저부 Meynert기저핵에서의 ChAT면역반응 신경세포들은 출생 후 발생과정에서 세포소기관과 신경돌기들의 분화에 따른 세포체 부피의 증가, 분화된 세포형들의 출현 율의 증가 및 세포의 손실이 없는 상태에서 세포 응축 등의 과정을 통하여 세포들이 분화한다고 생각된다.

• Applied Microscopy
• /
• 제28권2호
• /
• pp.139-158
• /
• 1998

The development of the superior cervical ganglion was studied by electron microscopic method in human fetuses ranging from 40 mm to 260 mm of crown-rump length (10 to 30 weeks of gestational age). At 40 mm fetus, the superior cervical ganglion was composed of clusters of undifferentiated cell, primitive neuroblast, primitive supporting cell, and unmyelinated fibers. At 70mm fetus, the neuroblasts and their processes were ensheated by the bodies or processes of satellite cells. The cytoplasm of the neuroblast contained rough endoplasmic reticulum, mitochondria, Golgi complex, Nissl bodies and dense-cored vesicles. As the neuroblasts grew and differentiated dense-cored vesicles moved away from perikaryal cytoplasm into developing processes. Synaptic contacts between the cholinergic axon and dendrites of postganglionic neuron and a few axosomatic synapses were first observed at 70 mm fetus. At 90 mm fetus the superior cervical ganglion consisted of neuroblasts, satellite cells, granule-containing cells, and unmyelinated nerve fibers. The ganglion cells increased somewhat in numbers and size by 150 mm fetus. Further differentiation resulted in the formation of young ganglion cells, whose cytoplasm was densely filled with cell organelles. During next prenatal stage up to 260 mm fetus, the cytoplasm of the ganglion cells contained except for large pigment granules, all intracytoplasmic structures which were also found in mature superior cervical ganglion. A great number of synaptic contact zones between the cholinergic preganglionic axon and the dendrites of the postganglionic neuron were observed and a few axosomatic synapses were also observed. Two morphological types of the granule-containing cells in the superior cervical ganglion were first identified at 90 mm fetus. Type I granule-containing cell occurred in solitary, whereas type II tended to appeared in clusters near the blood capillaries. Synaptic contacts were first found on the solitary granule-containing cell at 150 mm fetus. Synaptic contacts between the soma of type I granule-containing cells and preganglionic axon termials were observed. In addition, synaptic junctions between the processes of the granule- containing cells and dendrites of postganglionic neuron were also observed from 150 mm fetus onward. In conclusion, superior cervical ganglion cells and granule-containing cells arise from a common undifferentiated cell precursor of neural crest . The granule-containg cells exhibit a local modulatory feedback system in the superior cervical ganglion and nay serve as interneurons between the preganglionic and postganglionic cells.

• Tuberculosis and Respiratory Diseases
• /
• 제41권2호
• /
• pp.73-86
• /
• 1994

In addition to classic cholinergic and adrenergic pathways, the existence of a third division of autonomic control in the human airways has been proved. It is called a nonadrenergic noncholinergic(NANC) nervous system, and difficult to study in the absence of specific blockers. Neuropeptides are certainly suggested to be transmitters of this NANC nervous system. It is very frustrating to understand the pathophysiologic role of these peptides in the absence of any specific antagonists. However, further studies of neuropeptides might eventually lead to novel forms of treatment for bronchial asthma. Another study of the interaction between different components of the autonomic nervous system, either in ganglionic neurotransmission or by presynaptic modulation of neurotransmitters at the end-organ will elute neural control in airway disease, particularly in asthma. Studies of how autonomic control may be disordered in airway disease should lead to improvements in clinical management. Epithelial damage due to airway inflammation in asthma may induce bronchial hyperresponsiveness. Axon reflex mechanism is one of possible mechanisms in bronchial hyperresponsiveness. Epithelial damage may expose sensory nerve terminals and C-fiber nrve endings are stimulated by inflammatory mediators. Bi-directional communication between the nerves and mast cells may have important roles in allergic process. The psychological factors and conditioning of allergic reactions is suggested that mast cell activation might be partly regulated by the central nervous system via the peripheral nerves. Studies in animal models, in huamn airways in vitro and in patients with airway disease will uncover the interaction between allergic disease processes and psychologic factors or neural mechainsms.

• 한국산학기술학회논문지
• /
• 제18권1호
• /
• pp.295-301
• /
• 2017

보툴리눔 신경독소는 콜린성 신경말단(부)을 선택적으로 공격하여 신경마비를 일으키는 신경독소로서, 그람양성을 띠고 내성포자를 형성하는 절대혐기성 세균인 보툴리눔 균(Clostridium botulinum)이 만들어낸다. 이 중 보툴리눔 A형 독소(BoNT/A)는 음식물과 물을 오염시킬 수 있으며 생물 무기나 생물 협박물질로 사용될 수도 있다. 이 때문에 독성을 탐지할 수 있는 예민한 분석방법과 중독을 치료할 수 있는 효능 있는 항독소를 개발해낼 필요성이 제기되어 왔다. 본 연구에서는 BoNT/A를 중화할 수 있는 단일클론 항체(mAb)를 생산하기 위하여 BoNT/A로 면역된 토끼의 항혈청에서 유래한 scFv 라이브러리를 인간 IgG와 융합시켰다. 그렇게 재조합된 scFvIgG 항체 단백질을 안정된 세포주에서 발현시켰고 항체 친화 크로마토그래피를 사용하여 scFvIgG mAb 단백질을 정제하였다. ELISA로 정제된 scFvIgG mAb 단백질의 효율성을 확인하였고, in vivo 실험으로 BoNT/A에 대한 중화능을 시험하였다. 독성 중화능 실험은 마우스를 사용하여 수행하였는데, 그 결과 scFvIgG 항체(10 ug)는 BoNT/A(100,000 $LD_{50}$)의 독성이 주입된 마우스를 완전히 방어하지는 못하지만 마우스의 생존 기간을 현격하게 연장시키는 것이 확인되었다. 이러한 결과들은 이 scFvIgG mAb가 BoNT/A를 중화하는 효능을 가지고 있다는 점을 제시한다.