Ex. yeast (monocellular), chlamydomonus, plants, animals (multicellular)

Cell sizes

eukaryote (真核細胞)
    animal cell: 25 μm (average)
    young cell of seed plants: 5-25 μm
    red blood cell: 7 μm
    yeast: 6 μm

Major components of cells

protoplasm (原形質)

nuclear membrane (核膜)
nuclear sap (核液)
chromatin (染色質/糸) → chromosome (染色体)
nucleolus (核小体)

plasma membrane (形質膜)

cell membrane (細胞膜)

cytoplasm (細胞質)
mitochondria (ミトコンドリア)
ribosome (リボゾーム)
endoplasmic reticulum, ER (小胞体)

rough ER (粗面小胞体)
smooth ER (滑面小胞体) → microbody (ミクロボディ)

Golgi apparatus (ゴルジ体)
lysosome (リソゾーム)
central body (中心体)
chromoplast (色素体), in particular, chloroplast (葉緑体)
ribosome (リボゾーム)
microtubule (微小管)

metaplasm (後形質)

glycocalyx (糖衣) = polysaccharide + glycocacidic polysaccharide
→ having the abilities on water suction, ion exchange, and cell recognition

Function

• giving mechanical support
• resisting turgor pressure
• determining the growth and shape of the cell
• strengthening the plant
• influencing the apoplast transport
• due to its certain components serving as nutritive depot
• protecting the cell from mechanical and chemical influences
• yielding signal molecules for itself and for interacting organisms
• participating in cell-to-cell connections

Cell division

Middle lamella: present between cells and acts as a cement preventing cells from migrating

Components

• hemicellulose: alkaline-soluble
• pectinic substance
• cellulose
• protein
• lipid

Cell fusion (細胞融合)

Cell fusion of tomato (Lycopersicon esculentum Mill.) and potato (Solanum tuberosum L.)
supposedly succeed in conjugation culture of potato

The structure of plasma membrane is similar with that of tonoplast

the membreans are perviousness to water and oil → the mmembrean consists of the mosaic of lipids and proteins

the fats developed on the membrane surface of red blood cells consist of two molecules or sheets

Sandwich model of plasma membrane

repeating unit theory(反覆単位説)
peripheral nerve of frog → KMnO4, potassium per mangnite
unit membrane: electrons insde of the membrane do not scatter
→ inside: electron less dense / oudside: dense

Fig. Frog peripheral nerve

unit membrane (基本膜構造)

isolated the liver plasma membrane of rats by homogenization
membrane substances

fatty substrates + phospholipids + α-lecithin + β-lecithin = 40%
protein = 60%: most of them are structural proteins as enzymes
sugars: as glycoprotein in the plasma membrane

R1: (CH2)16CH3, R2: (CH2)7-CH=CH-CH2-CH=CH-(CH2)4-CH3

sialic acid(シアル酸) and N-acethylneuraminic acid (NANA) are focused, because these substances are considered to be related to electrical characteristics of membrane surfaces

phase transition between protein and lipid → transition between crystal and liquid-crystal conditions
→ denied later

negative staining of mitochondria by heavy metals → membrane consisting of protein-lipid complex

fluid mosaic theory (流動モザイク説)

FFMP (formyl methionyl methyl phosphate)
Characteristics:

• impermeability through cell membranes
• easily bonded with phospholipids (cell surface is bonded with glycoprotein and phospholipids)
• the presence inside or outside of membrane is known by radical phospholipids or not (indicating the bond with ³⁵S)

→ phosphatidyl ethanol amine is present in the inside of membrane
phospholyase A: enzyme hydrolyzing phospholipids
intact cell: 50% of total phospholipods are hydrolyzed

Phosphalidyl choline _________ 70-80% ___ foming outside
Sphingomyelin ______________ 70-80%
Phosphatidhyl ethanol-amine _____ 20% ___ forming inside
Phsophatidhyl serine ____________ 0%

the inside is also hydrolyzed when ghost cells are used

Outside __ SM ___ PC
Insdie ____ PE ___ PS

FMMP-³⁵S

intact red dry ghost: proteins → gel electrophoresis
gel electrophoresis of red cerp: intact red crep, ghost, proteins → gel electrophoresis
proteins etc. were separated by pore sizes → staining by commassie brilliant blue, amide black, etc.

    Band MW         Weight
    No   (dalton)   %
    1    24         15.7 ___ pectin
    2    21.5       14.7 ___ pectin
    2.1  20         -    ___ ankyrin
    3    8.8-8.9    24.1 ___ protein band #3
    4.1  7.8        4.2
    4.2  7.2        5.0
    5    4.3        4.5 ___ actin
    6    3.5        5.5
    7    2.9        3.4

Ex. much glycophorin (sugar) - commassie blue
60% sugar + 40% polypeptide (131 amino acids)
pAs staining (red) - pAs response
sugar chain is related to blood types and the field of binding with virus (influenza, hemagglutinating virus of Japan, etc.), and the field of recognizing lectin
pectin: penpheral protein extrinsic
ankyrin & band#3 protein: component A receptor
annnose 1: galactose 2: N-acetyl glucosamme 2
actin

Multiparticle model of cell membrane

chlorimecsterase

CH2-COR__________CH2OH
_|_______________→ |_______________+RCOOH + H2O
CH2-N⁺(CH3)3-OH^-___CH2N+(CH3)3OH^-

sugar → glucose + fructose

auxin(IAA)
dedol acetic acid

Selective permeability (選択的透過性)

Ghost (可逆溶血法): GTP, UTP, ITP - the permeability of red blood is increased when the blood is lowered into water
The transports of Na⁺ and K⁺ go across when ATP is supplied into the membrane

Giant axon(巨大軸索) of squid = giant nerve fiber
Na⁺ yield becomes low when cyan, azide, dinitrophenol(DNP) are given → oxidative phosphorylation
Na⁺ yield becomse high when ATP is supplied → ATP is required for sodium pump (Na pump)

○ K⁺ + Na⁺, ● lack K⁺ or Na⁺

ATPase → the membrane is activated region =
ATP → [Mg⁺⁺, Na⁺, K⁺] → ADP + Pi

1 molecule ATP
3Na⁺ atoms → pump out
2 K⁺ atoms → pump in

A-P-P-³²P + Na⁺ (in) + E₁(ATPase) →
[Mg⁺⁺] →
Na-E₁-³²P + ADP →
[K⁺, Mg⁺⁺] → Na-E₂-³²P →
K⁺ + H2O + Na-E2-³²P
⇔ E1 + Pi⁺
Na⁺ (outside), K⁺ (inside)

ATPase = 2.5-3.0 × 10⁵ = subunits (large 1.0-1.3 × 10⁵) + (small 0.5 × 10⁵)

couple (共載)
active transpport is performed by ATPase in the insdie of membrane

Endocytosis (エンドシトーシス, 飲細胞運動)): uptake

oragn or cell taking in nutrients → phagocytic vesicle

Exocytosis (エクソサイトーシス): evacuation

myofibrils enclosed by membrane structure

membrane structure within cells

Rough-surfaced endoplasmic reticulum, rER (粗面小胞体)

Smooth-surfaced endoplasmic reticulum, sER (滑面小胞体)

Microsomes (ミクロゾーム)

not present in viable and healthy cells

Centrosome (中心体)

Centriole (中心体)

1956 Bernband: structure → globe → 5 μm = diameter of tublin

sprial tube is formed by 13 tubulins are aggregated

present in flagellate (鞭毛虫) and ciliate (繊毛虫)
also found in Bryophytes and Pteridophytes
→ basal granule (keinetosomes, keineto nucleus (動原核), keinetoplast (毛基体)

9(3) + 2 → 9(3) + 3, 9(3) +1, 9(3) + 0

1968 Miki & Sakai (三木・酒井)

spindle fiber consisting of microbubules (confirmed by electron microscope)
Centrosome centriole

mitotic apparatus (紡錘体、中心体) → extracted from sea urchin and observed

Non-basic protein RNA Polysaccharide
______95%______6%_______+

Spindle fiber consisting mostly of proteins (without DNA) 5. Microtubules

1963 Slantherbach: cnidoblasts of hydra
1963 Ledbetter & Porter: Phleum pratense
Echinosphuycrium (太陽虫)

1952, 56 Loewy, Nakajima

proteins in Physarum are actin (because of mobility) – rejected in the present

1953, 67 Kitching

spirally lined-up microtubles → each microtuble consisting of 12 subunits?

1966 Nagai (永井), Rebhum: Chara

50-100 filaments of which diameter is 50Å
→ also discovered from Physarum in 1976

(called internal reticular apparatus) →
1988 the organella named after him in 1898

Function

cell division: mt ≈ bacteria

Handbook of histological and cytological technique. University of Chicago Press
vital staining by janus green → micro-granules are specifically stained
also stained by janus red, dahlia violet, toluidine blue, methylene blue

rat liver: 800 mitochondria/cell
human 500-2500/cell
sperm 10-25/cell

isolated mitochondria + Pi (リン酸) + ADP + NADH → ATP

increasing permeability by soaking into hypotonic solution
O2-aerobic

producing ATP → the measurements clarified (3 ATP)/NADH
required dicarbonic acids (ジカルボン酸) and tricarbonic acids (トリカルボン酸) for NADH synthesis
required O₂ for respiration

RC particles, supported fluid mosaic model

NAD: nicotic acid adenine dinucleotide (coenzyme I, DPN⁺): coenzyme
NADP: nicotic acid adenine dinucleotide phosphoric acid (coenzyme II, TPN⁺)
flavin mononucleotide (FMN)
Ubiquinone (CoQ, UQ)
→ oxidation

cytochrome oxidase lacked in mt ≠ the gene in nuclei

Structure

Function

C6H12O6 + O2 → 12H2O + 6CO2 + 38ATP (688 kcal)

Vesicles (小胞)

Chloroplast (葉緑体)

phaeoplast (褐色体) in brown algae and blue-green algae → fucoxanthin (褐藻素)
rhodoplast (紅色体) → phycoerythrin (紅色素)

Phylogeny

Spermatophyta: Grana-and intergrana-thylakoid
Pteridophyta, Bryophyta, Charophyta: Granoid (ferns), pseudo-grana (moss and Charophyta), or multiple thylakoid
Euglenophyta, Phaeophyta, Pyrrophyta, Chrysophyta: Triple thylakoid
Cryptophyta: Double thylakoid
Rhodophyta: Single thylakoid

Cyanophyta: Single or double thylakoid
Photosynthetic bacteria: Chromatophore or thylakoid

Structure

quantasome (etymology, quantum 光量) = ca 200 × 100 100 Å (80-100 Å)
contents in quantasome: protein, lipid, chlorophyll, carothin, cytochrome, etc.

regularly-arranged vesicles
developing prolamella body and primary lamella
growing defects in the darkness

growing for four days in the darkness → lamella structure developed from prolamella body rapidly after exposure to light

Stages: proplastid → differentiation → maturation

Proteins _______ 35-55 (half in lamella)

(Water soluble 20, Water insoluble 80)

Lipids _________ 20-30 (in lamella)
Carbohydrate ___ not constant (highly flctuated)
Pigments ______ 9 (as chlorophyll or kalotinoid)
Nucleic acids __ 2-3
+ NADP, NAD, FMN, FAD, ATP, ADP, NADPH₂ and NADH₂

☛ gene

Expression of gene information in chloroplasts

the concept of plastid continuity, stipulating that plastids do not arise de nove but from preexisting plastids (葉緑体有親説)

all cells are coming from smaller cells = all chloroplasts are comoing from the chloroplasts

Nitella → mutants: the cells can not divide cells when the chloroplants are lost
observed the chloroplast division is synchronized with the nucleus division

proposed proplastid-origin theory → the structures of immature mitochondria are similar with the structures of proplastids
zygote in embryonic cell (初期胚) →

considered that the chloroplasts are developed from the proplastids through deforming from immature mitochondria

Spirogyra, ³H-thymidine radio autography - confirming that chloroplast has DNA
high GC content for many plant species ⇔ DNA I: GC 60% = AT 40%, DNA II: GC 30% = AT 70%

Chloroplast – maternal inheritance (母性遺伝)

Chloroplast RNA

confirmed that RNA is present in chloroplast

¹⁴C-ATP: confirmed the synthesis of RNA polymerase

83S, 68S ⇒ sucrose density-gradient centrifugation - centrifuged high molecules

S: sedimentation coefficient (沈降定数), S-value, proposed by Sverdverg)
large S = large particles (high molar weight)

ribosome in leaf = 83S, 68S ↔ ribosome in root = only 83S
→ probably 68S = mitochondrial ribosomes → size ≈ bacterial ribosome
t-RNA: specific in chloroplast ↔ different from t-RNA in nucleus amino acids (a.a.) + ATP + enzyme → aminoacyl-AMP + PP

enzyme: aninoacyl-t-RNA synthetase

a.a.-AMP-Et t-RNA → aminoacyl-t-RNA
∴ Chloroplast RNA = Bacterial RNA
→ if the RNA is bacteria origin → RNAs and enzymes are differnt from them in cytoplasm

the presence of t-RNA and synthetase shown by adding ¹⁴C-valine and ATP

RNA containing ¹⁴C is synthesized when ¹⁴C-ATP, GTP, CTP and UTP are added - evidence on the synthesis
RNA synthetase and RNA polymerase

RNA extracted from the chloroplast is 3.25S

centrifugal separation of ribosomes from chloroplast just before and after exposing light
before: polysomes are absent ↔ after: present
add ¹⁴C-a.a. to isolated chloroplants → ¹⁴C is absorbed into the proteins in the chloroplasts

chlorophyll DNA: 40 μm
Assuming that the molecular weiht of polypeptides is 20000, the information in chloroplast DNA is for 100-200 genes
→ sufficient to cover all the enyzmatic responses

RNA polymerase is absent in chloroplasts
RNA polymerases are absent in the chloroplasts and are produced in the nuculei → same with DNA polymerase

Mitochondrial ribosome

cf. Yeast ribosome: 21S RNA + 15S RNA, Higher plants 26S + 18S

Hela's mitochondria → DNA, etc. → 16S RNA, 12S RNA
observed the RNAs after soaking into ferrum (Fe) by EM

Chloramphenicol – mitochondria origin (inhibiting protein synthesis)
Cycloheamoide – nucleus origin (inhibiting protein synthesis)

Nucleolus (仁)

observed that nucleoli remained in the three parts chromosome duirng mid-stage after heat treatment
→ secondary constriction was observed at the edge = probably nucleolus is distributed at the edge of chromosome
confirmed the similar locations were dyed by silver staining
→ conclusion: nucleolus is formed at specific locations in choromosome

Structure

two different structures in nucleolus - two patterns recognized by dye staining
vacuole (= nucleolus) or fibrous structure (= nucleolonema 仁糸)
inside: fibrons = RNA + proteins / outside: granular = ribosome-sized
→ nucleolus is considered to be precusor ribosomal particles condensed chromatin (=nucleolus associated chromatin): enclosed around nucleolus

Structural component

egg cells of starfish →
RNA = 3-5%, proteins = 85-95%, DNA = 0 (recently shown to be 0.2-10%)

Nuclear substances (核質)

chromatintin, nuclear sap, nucleolus, nuclear skeleton, nuclear vacuole

Chromatin (クロマチン)

discovered chromosomes in the dividing pollen mother cells of Tradescantia

described the splitting of dark stained rod-like structures during cell division which he called chromatin

discovered that chromosome number in a species remains constant

coined the term chromosome (chromo- = colored, -some = body)

Salivary gland chromosomes (唾腺染色体)

polytene chromosome (多糸染色体)

Lumpbrush chromosome (ランプブラシ染色体)

→ anucleate amebas cohabit

Structural and functional significance of bacteria and mitochondria
It has been, therefore, a constant source of encouragement and enlightenment for me to "rediscover" holistic concepts of mitochondria at a time when the observation of mitochondrial DNA fibrils was objected to, not on the basis of evidence presented, but because there was no obvious rationale for their presence. It was that, much to the dislike and must on occasion turn back in time to find original opinions on broad biological questions, because (a) our predecessors were here first, and (b) their brains were relatively uncultured to exercise their imagination.

proposed endosymbiosis (共生説)